Cd28 Mouse Monoclonal Antibody [Clone ID: JJ319]

Specifications

Product Data
Clone Name	JJ319
Applications	FC
Recommend Dilution	Flow Cytometry.
Reactivity	Rat
Host	Mouse
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	lot specific
Purification	Affinity chromatography on Protein G
Conjugation	FITC
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	Cd28 molecule
Database Link	Entrez Gene 25660 Rat UniProt ID P31042
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

*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.