Cd28 Mouse Monoclonal Antibody [Clone ID: JJ319]

CAT#: SM268BX

Cd28 mouse monoclonal antibody, clone JJ319, Biotin



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CNY 8,194.00


货期*
5周

规格
    • 500 ug

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Specifications

Product Data
Clone Name JJ319
Applications FC
Recommend Dilution Flow Cytometry (for details please see "Protocols" below).
Reactivity Rat
Host Mouse
Clonality Monoclonal
Immunogen Rat CD28 transfected A20J cells
Specificity Antibody SM268B recognizes a 90 kDa homeodimeric cell surface glycoprotein (CD28).
Results of tissue Distribution by Flow Cytometry Analysis:
Rat strain: Fischer
Cell concentration: 1x10e6 cells per test
Antibody concentration used: 1.0 µg/10e6 cells
Isotypic control: Biotin Mouse IgG1
Cell source percentage of cells stained above control
Thymus: 25.4%
Splenic T Cells*: 73.2%
*(T cells isolated with a Rat T Cell Recovery Column).
Formulation PBS containing 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
Label: Biotin
State: Liquid purified Ig fraction
Concentration lot specific
Purification Protein G Chromatography
Conjugation Biotin
Storage Condition Store at 2-8°C for up to one month. For longer storage, aliquot and freeze unused portion at -20°C in volumes appropriate for single usage.
Avoid freeze/thaw cycles.
Gene Name Cd28 molecule
Background CD28 has been found to be a potent costimulatory receptor on T cells. It is expressed on all peripheral rat alpha/beta and most gamma/delta T cells, as well as on approximately half of all NK cells.
Synonyms TP44
Note This clone can costimulate T cell proliferation and IL-2 secretion by resting rat T cells.

Protocol: FLOW CYTOMETRY ANALYSIS:
Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population.
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).
4. To each tube, add ~ 1.0 µg (optimize according to your assay conditions) of SM268B or SM268BX per 10e6 cells.
5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 µl of secondary antibody (Streptavidin-PE) at a appropriate dilution.
9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 µl ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

Media:
A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 µl of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 µl of 2M sodium azide in 100 mls).
Reference Data
*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.
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