Cd28 Mouse Monoclonal Antibody [Clone ID: JJ319]

CAT#: SM268FX

Cd28 mouse monoclonal antibody, clone JJ319, FITC


USD 650.00

2 Weeks*

Size
    • 500 ug

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Specifications

Product Data
Clone Name JJ319
Applications FC
Recommended Dilution Flow Cytometry.
Reactivities Rat
Host Mouse
Isotype IgG1
Clonality Monoclonal
Immunogen Rat CD28 transfected A20J cells
Donor: BALB/c spleen
Fusion Partner: X63-Ag 8.653
Specificity Anti-rat CD28 monoclonal antibody recognizes a 90kDa homeodimeric cell surface glycoprotein. CD28 has been found to be a potent costimulatory receptor on T cells. It is expressed on all peripheral rat ab and most gd T cells, as well as on approximately half of all NK cells.
This clone can costimulate T cell proliferation and IL-2 secretion by resting rat T cells.
Formulation PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
Label: FITC
State: Liquid purified Ig fraction
Label: Fluorescein isothiocyanate isomer 1
Concentration 0,1 mg/ml
Purification Affinity chromatography on Protein G
Conjugation FITC
Gene Name Rattus norvegicus Cd28 molecule (Cd28)
Synonyms TP44
Note Protocol: FLOW CYTOMETRY ANALYSIS:
Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-Rat cell separation medium.
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* of this antibody 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. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.)
7. Wash 2 times at 4°C.
8. Resuspend the cell pellet in 50 μl ice cold media B.
9. 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).
Results:
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: FITC Mouse IgG1
Percentage of cells stained above control:
Thymus 36.3%
Splenic T Cells* 79.7 %
*(T cells isolated with Rat T Cell Recovery Column Kit)
Reference Data

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*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.