MHC Class II (RT1D) antibody

Catalog No. ABIN114389

Product Details

Target: MHC Class II (RT1D)

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: Un-conjugated

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunoprecipitation (IP)

Purification: Protein G affinity chromatography

Immunogen: Rat spleen membrane glycoproteins depleted of Ia-A antigens Immunocyte Donor: BALB/c spleen Fusion Partner: X63 Ag8.653

Clone: OX-17

Isotype: IgG1

Application Details

Application Notes: Flow Cytometry: 1/250 - 1/500 (see Protocols). Immunhistochemistry. Immunoprecipitation.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: METHOD: 1. Prepare cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-Rat. cell separation medium. 2. Wash 2 times. 3. Resuspend cells to 1x10e6 cells in approximately 50 µl media A in a microcentrifugetube. (i. e. 50 µl of cells resuspended to 2x10e7 cells/ml). The contents of 1 tube represent 1test. 4. To each tube add 50 µl of a 1: 250 - 1: 500 dilution of this Ab. 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 (FITC Goat anti-mouse IgG (H+L)) at 1: 700 dilution. 9. Incubate tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in Media B. 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 propidiumiodide at 0. 5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalatingDNA). MEDIA: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2 M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% bovine serum albumin + sodium azide (100 µlof 2 M sodium azide in 100 mls). RESULTS: Rat Strain: Lewis RatCell Concentration: 1x10e6 cells per testAntibody Concentration: 1: 400Isotypic Control: Mouse IgG1, κCELL SOURCE PERCENT STAININGThymus 12%Spleen 34%Lymph Node 25%STRAIN DISTRIBUTION: Antibody Concentration: 1: 100Strains Tested: Wistar, Buffalo, Brown Norway, Fischer 344Positive: Wistar, Buffalo, Brown Norway, ACI, Fischer 344Negative: none

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS with 0.02 % sodium azide

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details

Target: MHC Class II (RT1D)

Alternative Name: MHC Class II RT1D

Background: Synonyms: HLA Class II