MHC Class II (RT1D) antibody (Biotin)

Catalog No. ABIN114385

This Mouse Monoclonal antibody specifically detects in FACS and IHC (fro). It exhibits reactivity toward Rat.

Quick Overview for MHC Class II (RT1D) antibody (Biotin) (ABIN114385)

Target: MHC Class II (RT1D)

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: Biotin

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

Clone: OX-17

Product Details

Purification: Protein G Chromatography

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

Isotype: IgG1

Application Details

Application Notes: Flow Cytometry. Immunohistochemistry on frozen sections.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a 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 the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 5-0. 1 µg* of this Ab 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-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom 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 propidiumiodide 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 (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Rat Strain: FischerCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 1 µg/10e6 cellsIsotypic Control: Biotin Mouse IgG1Cell Source Percentage of cells stained above control: Thymus 6. 1%Spleen 48. 8%Lymph Node 27. 5%

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL

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