MHC Class II I-Ab Mouse Monoclonal Antibody [Clone ID: 25-5-16S]

Specifications

Product Data
Clone Name	25-5-16S
Applications	CT, FC
Recommend Dilution	Flow cytometry: see protocol.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Specificity	This antibody specifically reacts with the I-Ab encoded MHC class II antigen expressed on mouse strains of the H-2b haplotype.
Formulation	PBS and 0.09% NaN3 State: Purified State: Liquid purified Ig fraction
Concentration	lot specific
Conjugation	Unconjugated
Storage Condition	Store the antibody at 2 - 8 °C up to one month or (in aliquots) at -20 °C for longer. Avoid repeated freezing and thawing.
Database Link	UniProt ID P18468
Background	Class II antigens are most highly expressed on antigen-presenting cells including B cells, macrophages, dendritic cells and certain epithelial cells.
Synonyms	H2-Eb1, H-2 class II histocompatibility antigen I-A beta chain
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x107 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1x106 cells, representing 1 test). 4. To each tube, add ~1.0 µg* of antibody. 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 IgM at 1:500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected 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 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). N.B Appropriate control samples should always be included in labelling studies. * For optimal results in various applications, it is recommended that each investigator determine dilutions appropriate for individual use.
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.