- Basic aspects of thoracic anaesthesia are general anesthesia often combined with regional anesthesia, intubation with double lumen tube and separation of lung ventilation. Proper assessment of pain and adequate analgesia in intraoperative and postoperative period is a challenging issue for medical practitioners.
- Intraoperative trauma may lead to many metabolic implications and disturbance of haemostasis, what can be reflected in change of blood and saliva hormone and other substance levels, such as alpha-amylase, cortisol, testosterone, secretory IgA, β-endorphin, nerve growth factor, calcitonin gene-related protein and P substance.
- The aim of this study is to assess the impact of regional anesthesia on hormone levels in postoperative period. Saliva was collected from participants in order to perform laboratory tests, using a special disposable Salivette tube (Sarstedt AG & Co, Germany). Saliva was collected by placing a sterile tampon under the tongue or chewing for 30-45 seconds.
- The soaked saliva pad was then placed in a suspended insert with a perforated bottom. The insert with a tampon was placed in a centrifuge tube and closed with a stopper. Next the tube was centrifuged (1000 x g for 10 min.) to obtain a ready to test saliva supernatant. Approximately 0,7 ml of the supernatant from every sample collected was used for further testing. Samples were frozen after centrifugation at – 85°C until performing laboratory tests. Blood was collected for laboratory tests from the ulnar vein. Blood for testing was collected using disposable equipment in a volume of 5ml into a tube containing ethylenediaminetetraacetic acid (EDTA) and aprotinin.
- Next the tube was centrifuged (1000 x g for 5 min.). After centrifugation and separation of Centrifuge tube sizes morphotic elements, the obtained plasma was divided into two tubes and frozen at – 85°C until performing laboratory tests.
Immune Responsiveness and Outcome After Aortic Valve Surgery (Measure)
- Barts Heart Centre is a 255-bedded specialist cardiac centre where approximately 200 primary isolated open aortic valve replacement procedures are performed per year. It is closely associated both geographically and academically with the William Harvey Research Institute (WHRI), which forms part of Queen Mary University, London (QMUL). This will allow laboratory analysis of all collected blood samples within 15 minutes if necessary. Barts Heart Centre is one of the largest cardiac units in Europe and a key research objective of QMUL/WHRI is to support cardiac research.
Sample collection and storage:
Following written informed consent all patients will be pre-operatively risk-scored by means of ASA, Euroscore 2 and conventionally risk-scored for the development of surgical site infection.
- Blood (44ml total) will be taken from 150 aortic valve replacement patients immediately prior to surgery (T0).
- Blood (7ml) will be collected into a PAXgene tube and stored at -80° for later genetic analysis. A clotted sample (7ml) will be collected in a standard gold top SST tube, left to clot then centrifuged for 10 mins at 1000g, the serum separated and stored at -80° for later analysis of antibody levels.
- In addition, 30 ml blood will be taken for study of peripheral blood mononuclear cells (PBMCs), drawn into standard purple top EDTA tubes (anticoagulant). These will be separated within 3 hours of collection by Ficoll-Paque density gradient centrifugation, whereby PBS-diluted blood is carefully layered over Ficoll and spun brake off in a centrifuge at 400g for 30 minutes.
- The PBMC layer is then extracted and washed twice in PBS and stored in 10% dimethyl sulfoxide freezing solution in liquid nitrogen so that they can be thawed, and batch analysed at a later date.
Sample analysis:
PBMCs will be counted using a haemocytometer, then cultured in RPMI and 10% serum with either LPS, α-toxin or unstimulated for 24 hours, after which they will be analysed immediately as detailed below.
• Soluble Mediators (Plasma)
– Storage: Ethylenediaminetetraacetic acid (EDTA, BD Biosciences Vacutainer, 9mL) anticoagulated blood will be centrifuged (1200g, 10mins, 20°C) and plasma stored at -80°C within 2hrs of collection
– Analysis: Cytokines: Meso Scale Discovery (MSD) V-PLEX Proinflammatory Panel 1 will be employed to quantify IFN-γ, IL-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6, IL-8, and TNF-α. Plates will be read using a MSD QuickPlex SQ 120 imager (Wiliam Harvey Institute, QMUL).
Antibodies: These will be measured by means of ELISA assays against staphylococcal antigens (α-toxin, teichoic acid) and core moieties of the endotoxin molecule (EndoCAb).
– Whole blood LPS/α-toxin Stimulated Cytokine Release
– Technique: As previously described and validated in our laboratory, heparinized blood will be stimulated for 4hrs (37°C, 250rpm) with 1ng/ml LPS within 2hrs of draw. After incubation, samples will be centrifuged (1200g, 10mins, 20°C) and supernatant stored at -80°C.
– Cytokine quantification: As per ‘soluble mediators’ MSD V-PLEX Proinflammatory Panel 1 will be employed to quantify TNF-α (primary outcome as metric of immune competence) and other cytokines. Cytokines will be expressed as a factor of the number of circulating monocytes.
Flow Cytometric Immunophenotyping and Functional Assessment of Leukocytes
Flow Cytometry: All samples will be analysed on the same machine (Becton Dickinson (BD) Fortessa, Rayne Building, UCL). Standardisation and comparison of output will be achieved via employment of constant voltages and compensation matrix throughout with daily matching of values to a single batch of Cytometry Setup and Tracking (CS&T) beads (BD). Leukocyte cell surface staining will be conducted using antibodies principally supplied by BD. Staining, data capture and storage will be conducted in accordance with a single study standard operating procedure. 5 panels (up to 14-colour) and two functional assays will be performed at each time-point. Data will be analysed in FlowJo version 10.5.
Surface Marker Staining:
Quantification and calibration panel: CD45, CD56, CD3, CD4, CD8, CD19, CD14, CD16, performed in a BD TruCount tube to enable enumeration of cell subsets Cell specific panels (incorporating markers of sub-categorisation and markers of immune competence)
– Monocyte/Myeloid: CD3, CD19, CD56, CD163, CD16, CD33, CD141, CD206, CD274, CD14, CD1c, CD11c, HLA-DR, CD80, CD123, CD83
– Neutrophil: CD3, CD19, CD56, HLA-DR, CD64, CD62L, CD11b, CD88, CD66b, CD14, CD11c, CD16, CD184, CD182
– Lymphocyte: CD19, CCR6, CD45RA, CD3, CD4, CD8, CXCR3, CD127, CD62L, CD25, CD56, CD27, CD279 HLA-DR Expression: Will be quantified on CD14+ monocytes using BD QuantiBrite beads.
Functional Measures Phagocytosis: The ability of neutrophils and monocytes to phagocytose opsonised FITC-labelled E.coli bacteria will be quantified via the PhagoTest assay (BD) as per manufacturer’s instructions Reactive oxygen ‘burst’: Quantitative determination of leukocyte oxidative burst will be undertaken using the PhagoBurst assay (BD) in response to un-labelled opsonized E.coli, phorbol 12-myristate 13-acetate (PMA) and the chemotactic peptide N-formyl-Met-Leu-Phe (fMLP).
The pre-surgery sample PBMCs that will be stimulated with LPS (type and concentration to be optimized via dose response curve on healthy volunteers) and α-toxin will undergo flow cytometry to measure TLR4 and TLR5 surface expression, HLA-DR surface expression and quantification of NF-kB using appropriate antibodies validated in the literature.
15 ml & 50 ml Centrifuge Tubes |
||
| TCF001-00-15mICentrifugeTubes450mlCentrifugeTubes4 | Vazyme | 15 mI Centrifuge Tubes*450 ml Centrifuge Tubes*4 |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Bag |
||
| LC8005-500 | GenDepot | 500/CS |
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Bag |
||
| LC8003-500 | GenDepot | 500/CS |
Centrifuge Tubes, 50 ml , Black ,PP,Sterile |
||
| LC8008-025 | GenDepot | 25/Bag |
Centrifuge Tubes, 50 ml , Black ,PP,Sterile |
||
| LC8008-500 | GenDepot | 500/CS |
Centrifuge Tubes, 50 ml , Amber ,PP,Sterile |
||
| LC8050-500 | GenDepot | 500/CS |
Centrifuge Tubes, 15 ml , Black ,PP,Sterile |
||
| LC8007-050 | GenDepot | 50/Bag |
Centrifuge Tubes, 15 ml , Black ,PP,Sterile |
||
| LC8007-500 | GenDepot | 500/CS |
Centrifuge Tubes, 15 ml , Amber ,PP,Sterile |
||
| LC8015-500 | GenDepot | 500/CS |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap,Self Standing, in Bag |
||
| LC8006-500 | GenDepot | 500/CS |
Centrifuge Tubes 0.65 ml Blue 10 bags of 1000 |
||
| MU11060 | Westburg | each |
Centrifuge Tubes 0.65 ml Pink 10 bags of 1000 |
||
| MU11120 | Westburg | each |
Centrifuge Tubes 1.7 ml Blue 10 bags of 500 |
||
| MU11430 | Westburg | each |
Centrifuge Tubes 1.7 ml Pink 10 bags of 500 |
||
| MU11490 | Westburg | each |
Centrifuge Tubes 2.0 ml Blue 10 bags of 400 |
||
| MU11920 | Westburg | each |
Centrifuge Tubes 2.0 ml Pink 10 bags of 400 |
||
| MU11980 | Westburg | each |
Rack S for centrifuge tubes 18 x 15 ml / 10 x 50 ml (45) |
||
| TP99017 | Westburg | each |
Rack L for centrifuge tubes 30 x 15 ml / 20 x 50 ml (30) |
||
| TP99019 | Westburg | each |
Centrifuge Tubes 0.65 ml Green 10 bags of 1000 |
||
| MU11080 | Westburg | each |
Centrifuge Tubes 1.7 ml Green 10 bags of 500 |
||
| MU11450 | Westburg | each |
Centrifuge Tubes 2.0 ml Green 10 bags of 400 |
||
| MU11940 | Westburg | each |
Centrifuge Tubes 0.65 ml Purple 10 bags of 1000 |
||
| MU11170 | Westburg | each |
Centrifuge Tubes 0.65 ml Yellow 10 bags of 1000 |
||
| MU11190 | Westburg | each |
Centrifuge Tubes 1.7 ml Yellow 10 bags of 500 |
||
| MU11560 | Westburg | each |
Centrifuge Tubes 2.0 ml Purple 10 bags of 400 |
||
| MU12020 | Westburg | each |
Centrifuge Tubes 2.0 ml Yellow 10 bags of 400 |
||
| MU12040 | Westburg | each |
50 mI Centrifuge Tubes |
||
| TCF00150-500Case | Vazyme | 500 / Case |
Centrifuge Tubes 0.65 ml Natural 10 bags of 1000 |
||
| MU11150 | Westburg | each |
Centrifuge Tubes 0.65 ml Rainbow 10 bags of 1000 |
||
| MU11210 | Westburg | each |
Centrifuge Tubes 1.7 ml Natural 10 bags of 500 |
||
| MU11510 | Westburg | each |
Centrifuge Tubes 1.7 ml Rainbow 10 bags of 500 |
||
| MU11590 | Westburg | each |
Centrifuge Tubes 2.0 ml Natural 10 bags of 400 |
||
| MU12000 | Westburg | each |
Centrifuge Tubes 2.0 ml Rainbow 10 bags of 400 |
||
| MU12060 | Westburg | each |
Centrifuge Tubes 0.65 ml Natural 20 bags of 1000 |
||
| MU17030 | Westburg | each |
Centrifuge Tubes 1.7 ml Natural 10 bags of 1000 |
||
| MU17040 | Westburg | each |
Centrifuge Tubes 2.0 ml Natural 10 bags of 1000 |
||
| MU17050 | Westburg | each |
15 mI Centrifuge Tubes |
||
| TCF00115-500Case | Vazyme | 500 / Case |
15 mL Centrifuge Tube, Tubes and Caps Packed Separately, Non-Sterile, 50/pk, 500 |
||
| 601151 | NEST Biotechnology | 500 pcs/cs |
Centrifuge tubes, glas, with round bottom, 50 ml |
||
| CD04-R50G | Phoenix instrument | each |
Centrifuge tubes, plastic, with round bottom, 50 ml |
||
| CD04-R50P | Phoenix instrument | each |
50ml Conical Centrifuge Tubes, Green Cap, 25/Bag |
||
| CT788-G | Bio Basic | 1PK, 25UNIT |
50ml Conical Centrifuge Tubes, Yellow Cap, 25/Bag |
||
| CT788-Y | Bio Basic | 1PK, 25UNIT |
Block, 15 x1.5ml centrifuge tubes |
||
| BCM1405-B15 | Bio Basic | 1 pcs, 1 UNIT |
Block, 15 x1.5ml centrifuge tubes |
||
| BSH100-15 | Benchmark Scientific | 1 PC |
Small centrifuge, 6 x 50 ml, 4000 rpm, incl. rotor for 6 x 50 ml or 6 x 15 ml Tubes |
||
| CD-0412-50 | Phoenix instrument | each |
Rack filled with 20 centrifuge tubes 50 ml, conical (10) |
||
| TP91550 | Westburg | each |
Stand For Centrifuge Tubes For 1.5 ml, |
||
| PW102-2NO | EWC Diagnostics | 1 unit |
Rack filled with 30 centrifuge tubes 15 ml, conical (10) |
||
| TP91515 | Westburg | each |
Centrifuge Tubes 0.65 ml, low binding 10x500 |
||
| MU11300 | Westburg | each |
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Rack |
||
| LC8004-500 | GenDepot | 500/CS |
Centrifuge Tubes 2.0 ml Natural 12 x 800 |
||
| MU1220 | Westburg | each |
Corning LSE Single Block 5 x 50 mL Centrifuge Tubes - EACH |
||
| 480129 | Scientific Laboratory Supplies | EACH |
29 mm Test Tube Foam Rack (for 50 mL centrifuge tubes) - EACH |
||
| MIX1118 | Scientific Laboratory Supplies | EACH |
Dolphin Centrifuge Tubes 2.0 ml Natural 5x1000 |
||
| MU16140 | Westburg | each |
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Rack |
||
| LC8002-500 | GenDepot | 500/CS |
16 mm Test Tube Foam Rack (for 15 mL centrifuge tubes) - EACH |
||
| MIX1098 | Scientific Laboratory Supplies | EACH |
Centrifuge Tubes 0.65 ml Natural Low binding 2x500 |
||
| MU11330 | Westburg | each |
Centrifuge Tubes 2.0 ml Natural, low binding 4x500 |
||
| MU12160 | Westburg | each |
Centrifuge Tubes 2.0 ml Natural Low binding 2x500 |
||
| MU12180 | Westburg | each |
Centrifuge Tubes 1.7 ml Natural, Low binding 5x500 |
||
| MU39640T | Westburg | each |
15 mL Centrifuge Tube Rack, 1/pk, 50/cs |
||
| 610001 | NEST Biotechnology | 50 pcs/cs |
Centrifuge tube 50 ml, conical / triple bagged (50) |
||
| TP391050 | Westburg | each |
Centrifuge Tubes 1.7 ml Natural Lubricated 2 x 500 |
||
| MU11720 | Westburg | each |
4-Ways Microtube and Centrifuge Tubes Racks, Red |
||
| R023-R | Bio Basic | 1 UNIT |
4-Ways Microtube and Centrifuge Tubes Racks, Blue |
||
| R023-B | Bio Basic | 1 UNIT |
Greiner centrifuge tubes - PK300 |
||
| T2193-300EA | Scientific Laboratory Supplies | PK300 |
Greiner centrifuge tubes - PK1000 |
||
| T1943-1000EA | Scientific Laboratory Supplies | PK1000 |
4-Ways Microtube and Centrifuge Tubes Racks, Yellow |
||
| R023-Y | Bio Basic | 1 UNIT |
4-Ways Microtube and Centrifuge Tubes Racks, Natural |
||
| R023-N | Bio Basic | 1 UNIT |
Centrifuge tube 50 ml, round (180) |
||
| TP91056 | Westburg | each |
Block, 15 x 1.5ml or 2.0ml centrifuge tubes (conical) |
||
| BCM1405-B1520 | Bio Basic | 1 pcs, 1 UNIT |
Block, 15 x 1.5ml or 2.0ml centrifuge tubes (conical) |
||
| BSH100-1520 | Benchmark Scientific | 1 PC |
Centrifuge Tubes, Autoclavable BLUE |
||
| CG264-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge Tubes, Autoclavable Round |
||
| CG276-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge Tubes, Autoclavable Round |
||
| CG277-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge Tubes, Autoclavable Round |
||
| CG278-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge Tubes, Autoclavable Round |
||
| CG279-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge tube 50 ml, conical (360) |
||
| TP91050 | Westburg | each |
Centrifuge tube 50 ml, conical (160) |
||
| TP91054 | Westburg | each |
Block for 5ml centrifuge tubes |
||
| H5000-5MT | Benchmark Scientific | 1 PC |
15 mL Centrifuge Tube, Bulk, Sterile, 50/pk, 500/cs |
||
| 601052 | NEST Biotechnology | 500 pcs/cs |
Centrifuge Tubes, Autoclavable Conical |
||
| CG274-100X1NO | EWC Diagnostics | 1 unit |
Centrifuge tube 15 ml, conical (360) |
||
| TP91014 | Westburg | each |
Centrifuge tube 15 ml, conical (800) |
||
| TP91015 | Westburg | each |
Centrifuge tube 15 ml, conical (800) |
||
| TP91115 | Westburg | each |
15mL Sterile Centrifuge Tubes PP |
||
| ARG1818 | Scientific Laboratory Supplies | PK500 |
50mL Sterile Centrifuge Tubes PP |
||
| ARG1824 | Scientific Laboratory Supplies | PK500 |
Block, 20x10mm test tubes or 20x2.0ml centrifuge tubes |
||
| BCM1412-BSW10 | Bio Basic | 1 pcs, 1 UNIT |
Block, 20x10mm test tubes or 20x2.0ml centrifuge tubes |
||
| BSW10 | Benchmark Scientific | 1 PC |
Eppendorf Adapter for 0.5 mL micro- centrifuge tubes + 0.6 ml Microtainer set of 6 - PK6 |
||
| E5425716001 | Scientific Laboratory Supplies | PK6 |
CapLock 0.5mL Centrifuge Tubes - PK500 |
||
| ARG1782 | Scientific Laboratory Supplies | PK500 |
Centrifuge tube 15 ml, conical / triple bagged (100) |
||
| TP391015 | Westburg | each |
Oakridge Centrifuge Tubes 30ml - PK10 |
||
| 31390030 | Scientific Laboratory Supplies | PK10 |
CapLock 1.5mL Centrifuge Tubes - PK1000 |
||
| ARG1808 | Scientific Laboratory Supplies | PK1000 |
Block, 24 x 0.5ml centrifuge tubes |
||
| BCM1405-B05 | Bio Basic | 1 pcs, 1 UNIT |
Block, 24 x 0.5ml centrifuge tubes |
||
| BCM1412-BSW05 | Bio Basic | 1 pcs, 1 UNIT |
Block, 24 x 0.5ml centrifuge tubes |
||
| BSW05 | Benchmark Scientific | 1 PC |
Block, 24 x 0.5ml centrifuge tubes |
||
| BSH100-05 | Benchmark Scientific | 1 PC |
50 mL Centrifuge Tube Rack, 1/pk, 50/cs |
||
| 610101 | NEST Biotechnology | 50 pcs/cs |
Centrifuge Tubes S 2420 30ml Qfit - PK10 |
||
| QBC24C24R0 | Scientific Laboratory Supplies | PK10 |
Centrifuge Tubes, Autoclavable RED Cap, |
||
| CG272-1X100NO | EWC Diagnostics | 1 unit |
Block, 4 x 15ml centrifuge tubes |
||
| BCM1405-B150 | Bio Basic | 1 pcs, 1 UNIT |
Block, 2 x 50ml centrifuge tubes |
||
| BCM1405-B500 | Bio Basic | 1 pcs, 1 UNIT |
Block, 12 x 15ml centrifuge tubes |
||
| BCM1412-BSW15 | Bio Basic | 1 pcs, 1 UNIT |
Block, 5 x 50ml centrifuge tubes |
||
| BCM1412-BSW50 | Bio Basic | 1 pcs, 1 UNIT |
Block, 4 x 15ml centrifuge tubes |
||
| BSH100-150 | Benchmark Scientific | 1 PC |
Block, 2 x 50ml centrifuge tubes |
||
| BSH100-500 | Benchmark Scientific | 1 PC |
Block, 12 x 15ml centrifuge tubes |
||
| BSW15 | Benchmark Scientific | 1 PC |
Block, 5 x 50ml centrifuge tubes |
||
| BSW50 | Benchmark Scientific | 1 PC |
Centrifuge Tubes, with ring (leak proof) |
||
| PW1207-1X100NO | EWC Diagnostics | 1 unit |
Centrifuge Tubes, Autoclavable With RED |
||
| CG271-1X100NO | EWC Diagnostics | 1 unit |
Block for 5mL centrifuge tubes - EACH |
||
| MIX7218 | Scientific Laboratory Supplies | EACH |
Block, 24 x1.5ml centrifuge tubes (conical) |
||
| BCM1412-BSW1500 | Bio Basic | 1 pcs, 1 UNIT |
Block, 24 x1.5ml centrifuge tubes (conical) |
||
| BSW1500 | Benchmark Scientific | 1 PC |
15mL Sterile Centrifuge Tubes PP - PK500 |
||
| ARG1814 | Scientific Laboratory Supplies | PK500 |
50mL Sterile Centrifuge Tubes PP - PK500 |
||
| ARG1820 | Scientific Laboratory Supplies | PK500 |
Falcon Conical 50ml PP Centrifuge Tubes Bagged - PK500 |
||
| 352070 | Scientific Laboratory Supplies | PK500 |
Falcon Conical 15ml PP Centrifuge Tubes Bagged - PK500 |
||
| 352096 | Scientific Laboratory Supplies | PK500 |
50ml Conical Centrifuge Tubes, Polypropylene |
||
| 21-106 | Genesee Scientific | 20 Racks of 25 Tubes, 500/Unit |
50ml Conical Centrifuge Tubes, Polypropylene |
||
| 21-108 | Genesee Scientific | 20 Bags of 25 Tubes, 500/Unit |
15ml Conical Centrifuge Tubes, Polypropylene |
||
| 28-101 | Genesee Scientific | 20 Racks of 25 Tubes, 500/Unit |
15ml Conical Centrifuge Tubes, Polypropylene |
||
| 28-103 | Genesee Scientific | 20 Bags of 25 Tubes, 500/Unit |
50ml Conical Centrifuge Tubes, Polypropylene |
||
| 28-106 | Genesee Scientific | 20 Racks of 25 Tubes, 500/Unit |
50ml Conical Centrifuge Tubes, Polypropylene |
||
| 28-108 | Genesee Scientific | 20 Bags of 25 Tubes, 500/Unit |
50ml Plug-Seal Centrifuge Tubes with Rack, 25 tubes/rack |
||
| 10-0501 | IHC World | - |
Kimas high strength centrifuge tubes - PK6 |
||
| Z514861-6EA | Scientific Laboratory Supplies | PK6 |
5 mL Centrifuge Tubes, Clear, Round, Lock Cap, Sterile, 200/pk, 2000/cs |
||
| 603111 | NEST Biotechnology | 2000 pcs/cs |
Centrifuge 5804 + Rotor S-4-72 + adapters 15/50ml tubes - EACH |
||
| E5804000565 | Scientific Laboratory Supplies | EACH |
Tubes Centrifuge 15ml Conical - PK500 |
||
| CEN9202 | Scientific Laboratory Supplies | PK500 |
15 mL Centrifuge Tube, Rack(PS Rack), Sterile, 50/pk, 500/cs |
||
| 601051 | NEST Biotechnology | 500 pcs/cs |
CENTRIFUGE TUBE 50 ML 30 X 115 MM - PK500 |
||
| T2318-500EA | Scientific Laboratory Supplies | PK500 |
Centrifuge 5804R + Rotor S-4-72 + adapters 15/50ml tubes - EACH |
||
| E5805000665 | Scientific Laboratory Supplies | EACH |
Centrifuge 5810 + Rotor S-4-104 + adapters 15/50ml tubes - EACH |
||
| E5810000560 | Scientific Laboratory Supplies | EACH |
250 mL PP Centrifuge Tubes with Plug Seal Cap, Sterile, 6/pk, 102/cs |
||
| 622001 | NEST Biotechnology | 102 pcs/cs |
500 mL PP Centrifuge Tubes with Plug Seal Cap, Sterile, 6/pk, 36/cs |
||
| 623001 | NEST Biotechnology | 36 pcs/cs |
Centrifuge 5810R + Rotor S-4-104 + adapters 15/50ml tubes - EACH |
||
| E5811000660 | Scientific Laboratory Supplies | EACH |
SINGLE BLOCK, 12 X 15ML CENTRIFUGE TUBES |
||
| 480124 | CORNING | 1/pk |
Centrifuge tube 50 ml, conical, with rim (320) |
||
| TP91051 | Westburg | each |
Block, 24 x 1.5ml or 2.0ml centrifuge tubes |
||
| BCM1412-BSW1520 | Bio Basic | 1 pcs, 1 UNIT |
Block, 24 x 1.5ml or 2.0ml centrifuge tubes |
||
| BSW1520 | Benchmark Scientific | 1 PC |
250mL centrifuge tube PP (59.8 X 160mm), with screw-cap, 14 bags of 5 tubes |
||
| C1250 | MTC Bio | 70 |
Block, 15 x 1.5ml Centrifuge Tubes for Dry Bath with Heated Lid |
||
| BCM1405-B15HL | Bio Basic | 1 pcs, 1 UNIT |
Block, 15 x 1.5ml Centrifuge Tubes for Dry Bath with Heated Lid |
||
| BSH100-15HL | Benchmark Scientific | 1 PC |
Falcon Conical 225ml PP Centrifuge Tubes - PK48 |
||
| 352075 | Scientific Laboratory Supplies | PK48 |
5 mL Centrifuge Tubes, Clear, Round, Lock Cap, Non-Sterile, 200/pk, 2000/cs |
||
| 603011 | NEST Biotechnology | 2000 pcs/cs |
15mL and 50ml Universal Rack for Conical Bottom Centrifuge Tubes - PK20 |
||
| 4366 | Scientific Laboratory Supplies | PK20 |
Falcon Conical 15ml Ps Centrifuge Tubes - PK500 |
||
| 352099 | Scientific Laboratory Supplies | PK500 |
5-Well Centrifuge Tube Box (1.5ml To 2ml Tubes) |
||
| R005-G | Bio Basic | 1 UNIT |
High Speed Centrifuge, Adapt for 1.5/2.0 mL Microcentrifuge Tubes, 500-1200 RPM |
||
| 102101 | NEST Biotechnology | 1 pcs/cs |
250 mL PP Centrifuge Tubes with Plug Seal Cap, Racked, Sterile, 6/pk, 24/cs |
||
| 622002 | NEST Biotechnology | 24 pcs/cs |
500 mL PP Centrifuge Tubes with Plug Seal Cap, Racked, Sterile, 6/pk, 24/cs |
||
| 623002 | NEST Biotechnology | 24 pcs/cs |
Block, 6 x 5ml centrifuge tubes (17mm diameter) |
||
| BCM1405-B5MT | Bio Basic | 1 pcs, 1 UNIT |
Block, 12 x 5.0ml centrifuge tubes (17mm diameter) |
||
| BCM1412-BSW5MT | Bio Basic | 1 pcs, 1 UNIT |
Block, 6 x 5ml centrifuge tubes (17mm diameter) |
||
| BSH100-5MT | Benchmark Scientific | 1 PC |
Block, 12 x 5.0ml centrifuge tubes (17mm diameter) |
||
| BSW5MT | Benchmark Scientific | 1 PC |
Block, 8x5ml centrifuge tubes, for MultiTherm Touch |
||
| H5100-5MT | Benchmark Scientific | 1 each |
Block 15 x 1.5mL Centrifuge Tubes for Dry Bath with Heated Lid - EACH |
||
| BLO1254 | Scientific Laboratory Supplies | EACH |
Centrifuge 5804 + Rotor S-4-72 + adapters 15/50ml tubes- IVD Only - EACH |
||
| E5804000560 | Scientific Laboratory Supplies | EACH |
Centrifuge Tube, 5ml, 200 per. pk. (4 bags of 50) |
||
| C1005-T5-Clear | Benchmark Scientific | 1 each |
Centrifuge Tube, 5ml, 200 per. pk. (4 bags of 50) |
||
| C1005-T5_Clear | Benchmark Scientific | each |
Centrifuge 5804R + Rotor S-4-72 + adapters 15/50ml tubes- IVD Only - EACH |
||
| E5805000661 | Scientific Laboratory Supplies | EACH |
Centrifuge 5810 + Rotor S-4-104 + adapters 15/50ml tubes- IVD Only - EACH |
||
| E5810000564 | Scientific Laboratory Supplies | EACH |
15ml Centrifuge Tube, Screw Cap, Natural, 100/Bag |
||
| BT888-N | Bio Basic | 1PK, 100UNIT |
Centrifuge 5810R + Rotor S-4-104 + adapters 15/50ml tubes- IVD Only - EACH |
||
| E5811000665 | Scientific Laboratory Supplies | EACH |
50mL Rack for Conical Bottom Centrifuge Tubes - PK20 |
||
| 4365 | Scientific Laboratory Supplies | PK20 |
15mL Rack for Conical Bottom Centrifuge Tubes - PK20 |
||
| 431355 | Scientific Laboratory Supplies | PK20 |
50 mL Centrifuge Tube, Bulk, Sterile, 25/pk, 500/cs |
||
| 602052 | NEST Biotechnology | 500 pcs/cs |
Falcon Conical 175ml PP Centrifuge Tubes Racked - PK48 |
||
| 352076 | Scientific Laboratory Supplies | PK48 |
Cool rack 9 wells for 15ml centrifuge tubes - EACH |
||
| HOM2740 | Scientific Laboratory Supplies | EACH |
Cool rack 4 wells for 50ml centrifuge tubes - EACH |
||
| HOM2742 | Scientific Laboratory Supplies | EACH |
Falcon Conical 15ml PP Centrifuge Tubes Racked - PK500 |
||
| 352097 | Scientific Laboratory Supplies | PK500 |
Falcon Conical 50ml PP Centrifuge Tubes Racked - PK500 |
||
| 352098 | Scientific Laboratory Supplies | PK500 |
Kimax Reuseable Glass Centrifuge tubes oil 203mm - PK12 |
||
| TUB2716 | Scientific Laboratory Supplies | PK12 |
15 mL Centrifuge Tube, Amber Color, Rack(PS Rack), Sterile, 50/pk, 500/cs |
||
| 601201 | NEST Biotechnology | 500 pcs/cs |
Block 6 x 5mL centrifuge tubes (17mm diameter) - EACH |
||
| BLO1256 | Scientific Laboratory Supplies | EACH |
Block 12 x 5.0mL centrifuge tubes (17mm diameter) - EACH |
||
| BLO1276 | Scientific Laboratory Supplies | EACH |
50ml Skirted Centrifuge Tube, Green Cap, 25/Bag |
||
| TK943-G | Bio Basic | 1PK, 25UNIT |
50ml Skirted Centrifuge Tube, Yellow Cap, 25/Bag |
||
| TK943-Y | Bio Basic | 1PK, 25UNIT |
Falcon Conical 15ml Ps Multipack Centrifuge Tubes - PK500 |
||
| 352095 | Scientific Laboratory Supplies | PK500 |
15ml Centrifuge Tube, Screw Cap, Red, Sterile, 100/Bag |
||
| BT888-SR | Bio Basic | 1PK, 100UNIT |
Centrifuge tube 13 ml, flat (800) |
||
| TP91019 | Westburg | each |
15ml Centrifuge Tube, Screw Cap, Blue, Sterile, 100/Bag |
||
| BT888-SB | Bio Basic | 1PK, 100UNIT |
CoolRack XT 5ml holds 12 x 5ml centrifuge tubes - EACH |
||
| 432060 | Scientific Laboratory Supplies | EACH |
15mL Flip Top Centrifuge Tubes Sterile DNase Free - PK500 |
||
| ARG1154 | Scientific Laboratory Supplies | PK500 |
50mL Flip Top Centrifuge Tubes Sterile DNase Free - PK500 |
||
| ARG1156 | Scientific Laboratory Supplies | PK500 |
50 ml Centrifuge Tube with Flat Cap, Bulk, Sterile (20x25) |
||
| OLOCT-50B | Westburg | each |
Snap Cap Polyethylene For Centrifuge Tubes Size 2 - PK500 |
||
| TUB2728 | Scientific Laboratory Supplies | PK500 |
15ml Centrifuge Tube, Screw Cap, Yellow, Sterile, 100/Bag |
||
| BT888-SY | Bio Basic | 1PK, 100UNIT |
Benchmark Centrifuge Tube Red 5mL (4 bags of 50) - PK200 |
||
| CEN2048 | Scientific Laboratory Supplies | PK200 |
15 ml Centrifuge Tube with Flat Cap, Bulk, Sterile (20x25) |
||
| OLOCT-15B | Westburg | each |
15ml Centrifuge Tube, Screw Cap, Natural, Sterile, 100/Bag |
||
| BT888-SN | Bio Basic | 1PK, 100UNIT |
50 ml Centrifuge Tube with Flat Cap, Racked, Sterile (20x25) |
||
| OLOCT-50R | Westburg | each |
Benchmark Centrifuge Tube Blue 5mL (4 bags of 50) - PK200 |
||
| CEN2046 | Scientific Laboratory Supplies | PK200 |
Benchmark Centrifuge Tube Clear 5mL (4 bags of 50) - PK200 |
||
| CEN2045 | Scientific Laboratory Supplies | PK200 |
Benchmark Centrifuge Tube Green 5mL (4 bags of 50) - PK200 |
||
| CEN2047 | Scientific Laboratory Supplies | PK200 |
Benchmark Centrifuge Tube Black 5mL (4 bags of 50) - PK200 |
||
| CEN2051 | Scientific Laboratory Supplies | PK200 |
50ml High Speed Centrifuge Tube, Natural cap, 25/Bag |
||
| CT790-N | Bio Basic | 1PK, 25UNIT |
15 ml Centrifuge Tube with Flat Cap, Racked, Sterile (20x25) |
||
| OLOCT-15R | Westburg | each |
Benchmark Centrifuge Tube Yellow 5mL (4 bags of 50) - PK200 |
||
| CEN2049 | Scientific Laboratory Supplies | PK200 |
50 mL Centrifuge Tube, Rack (PS Rack), Sterile, 25/pk, 500/cs |
||
| 602051 | NEST Biotechnology | 500 pcs/cs |
50ml Skirted Centrifuge Tube, Green Cap, Sterile, 25/Bag |
||
| TK943-GS | Bio Basic | 1PK, 25UNIT |
Centrifuge tube 16 ml, round, long (800) |
||
| TP91017 | Westburg | each |
50ml Skirted Centrifuge Tube, Yellow Cap, Sterile, 25/Bag |
||
| TK943-YS | Bio Basic | 1PK, 25UNIT |
Centrifuge tube 13 ml, round, short (800) |
||
| TP91016 | Westburg | each |
Corning 15mL PP Centrifuge Tubes Rack Packed with CentriStar Cap Sterile 50/Rack 500/Case - PK500 |
||
| 430790 | Scientific Laboratory Supplies | PK500 |
15ml centrifuge tubes, rack pack, sterile, flat top caps, graduation |
||
| SGB011-R | SEEN GROUP LIMITED | 50pcs/rack, 500pcs/carton |
50ml centrifuge tubes, rack pack, sterile, flat top caps, graduation |
||
| SGB012-R | SEEN GROUP LIMITED | 25pcs/rack, 500pcs/carton |
Kimax Centrifuge tubes graduated beaded top white scale 10ml - PK12 |
||
| TUB2708 | Scientific Laboratory Supplies | PK12 |
DB 2.1 Single block for 12 x 15ml conical centrifuge tubes - EACH |
||
| BLO6032 | Scientific Laboratory Supplies | EACH |
50 mL Centrifuge Tube, Self-Standing, Bulk, Sterile, 25/pk, 500/cs |
||
| 602072 | NEST Biotechnology | 500 pcs/cs |
Kimax Reuseable Glass Centrifuge tubes plain high strength 15ml - PK6 |
||
| TUB2718 | Scientific Laboratory Supplies | PK6 |
50ml High Speed Centrifuge Tube, Natural cap, Sterile, 25/Bag |
||
| CT790-SN | Bio Basic | 1PK, 25UNIT |
MICRO CENTRIFUGE TUBE, AMBER, 5 ML, 250/1250 |
||
| MCT-500-X | CORNING | 250/pk |
50ml Conical Centrifuge Tubes, Green Cap, Sterile, 25unit (cardboard rack) |
||
| CT788-GS | Bio Basic | 1PK, 25UNIT |
50ml Conical Centrifuge Tubes, Yellow Cap, Sterile, 25unit (cardboard rack) |
||
| CT788-YS | Bio Basic | 1PK, 25UNIT |
Holder for up to eight 15ml centrifuge tubes, for Bench-Top Shaker |
||
| 099ASR815 | Glascol | each |
At the end of stimulation, cells will be harvested, and RNA extracted for quantitative (q)PCR to evaluate AID mRNA expression. Although B cells in the PBMC cultures have been stimulated in the presence of other cell types, primarily T cells and monocytes-macrophages, our endpoint is to measure a B-cell response, as AID is exclusively expressed in B cells.