MHC Class II I-A antibody

Catalog No. ABIN114304

Product Details anti-MHC Class II I-A Antibody

Target: MHC Class II I-A

Reactivity: Mouse, Human

Host: Mouse

Clonality: Polyclonal

Conjugate: This MHC Class II I-A antibody is un-conjugated

Application: Cytotoxicity Test (CyTox)

Purification: Alloantiserum

Application Details

Application Notes: As a cytotoxic antibody, it can be used with complement for the enumeration orelimination of B cells, and is particularly well-suited as a positive control serum for B celltyping. The antiserum can also be used for biochemical studies of HLA-DR molecules.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: METHOD FOR USE WITH MOUSE LYMPHOCYTES: RECOMMENDED METHOD FOR DEPLETING A MOUSE CELL POPULATION OF Ia ANTIGENBEARING CELLS: 1. Prepare a cell suspension from the appropriate tissue (e. g. spleen, lymph node, etc. ) inCytotoxicity Medium1 or equivalent. Remove erythrocytes and dead cells (where necessary)by purification on Lympholyte®-M density cell separation medium2. After washing, adjustthe cell concentration to 1. 1x10e7 cells per ml in cytotoxicity medium. 2. Add the anti-Ia antiserum to a final concentration of 1: 20. 3. Incubate for 60 minutes at 4°C. 4. Centrifuge to pellet the cells and discard the supernatant. 5. Resuspend to the original volume in cytotoxicity medium containing the appropriateconcentration of Low-Tox®-M Rabbit Complement3,4. 6. Incubate for 60 minutes at 37°C. 7. Place on ice and monitor for percent cytotoxicity before further processing. For thispurpose, remove a small sample from each tube, dilute 1: 10, and add 1/10 volume of 1%Trypan Blue. After 3-5 minutes, score live versus dead cells in a hemacytometer. 8. For functional studies, remove dead cells from treated groups before further processing,particularly if the treated cells are to be cultured. Layering the treated cell suspension overan equal volume of Lympholyte-M cell separation medium and centrifuging, as per theinstructions provided, can do this. Live cells will form a layer at the interface, while thedead cells pellet. The interface can then be collected in cytotoxicity medium before beingresuspended in the appropriate medium for further processing. RECOMMENDED METHOD FOR DETERMINING PERCENT OF Ia ANTIGEN BEARING CELLS IN AMOUSE CELL POPULATION: 1. Prepare a cell suspension from the appropriate tissue in Cytotoxicity Medium orequivalent. Remove erythrocytes and dead cells (where necessary) by purification onLympholyte®-M density cell separation medium2. After washing, adjust the cellconcentration to 1. 1x10e6 cells per ml in cytotoxicity medium. 2. Add the anti-Ia antiserum to a final concentration of 1: 60.

Restrictions: For Research Use only

Handling

Format: Liquid

Reconstitution: Restore with 1 mL of cold distilled water.

Handling Advice: This product is photosensitive and should be protected from light

Storage: 4 °C

Storage Comment: Prior to and following reconstitution store the antibody undiluted at 2-8 °C. DO NOT FREEZE!

Target Details for MHC Class II I-A

Target: MHC Class II I-A

Alternative Name: MHC Class II Ia (MHC Class II I-A Products)