ab25228 has been shown to block H2Ld-specific and H2Ld-restricted antigen-specific lysis of target cells by cytotoxic T lymphocytes, but it does not block recognition of H2Ld-positive target cells by Ly-6G2-positive NK cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Primary antibody notesab25228 has been shown to block H2Ld-specific and H2Ld-restricted antigen-specific lysis of target cells by cytotoxic T lymphocytes, but it does not block recognition of H2Ld-positive target cells by Ly-6G2-positive NK cells.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use a concentration of 2 µg/ml.
Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
RelevanceIn the mouse the MHC Class 1 loci are called H2K, H2L and H2D. The equivalent loci in the human MHC are HLA (Human Leukocyte Antigen).
MHC class I molecules are heterodimers, consisting of a single transmembrane polypeptide chain (the a-chain) which is highly polymorphic, and the invariant ß2 microglobulin (which is encoded elsewhere, not in the MHC). MHC class I molecules are found on almost every nucleated cell of the body. Their major function is to present peptide fragments derived from antigens to cytotoxic T cells.
Cellular localizationCell Membrane; single pass type I membrane protein
H2 class I histocompatibility antigen, DB alpha chain antibody
Histocompatibility 2, D region locus 1 antibody
Anti-MHC Class 1 H2 Db antibody [28-14-8] images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class 1 H2 Db antibody [28-14-8] (ab25228)
IHC image of MHC Class 1 H2 Db staining in mouse spleen formalin fixed paraffin embedded tissue section, using Mouse-on-Mouse detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 20 mins. The section was incubated with ab25228, 10µg/ml, for 15 mins at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
Flow Cytometry - Anti-MHC Class 1 H2 Db antibody [28-14-8] (ab25228)
Flow Cytometry analysis of BALB/c mouse splenocytes labeling MHC Class 1 H2 Db with ab25228 at 1 μg/106 cells (purple). A Goat Anti-Mouse IgG2a, Human ads-FITC was used as the secondary antibody. Grey - Isotype Control, Mouse IgG2a-UNLB followed by Goat Anti-Mouse IgG2a, Human ads-FITC.
Flow Cytometry-MHC Class 1 H2 Db antibody [28-14-8](ab25228)
Overlay histogram showing SP0/2 cells stained with ab25228 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab25228, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
References for Anti-MHC Class 1 H2 Db antibody [28-14-8] (ab25228)