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Anti-HLA Antibody EP1395Y
|A synthetic peptide corresponding to residues in human MHC class 1 was used as an immunogen|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||IHC-Fr: Use at an assay dependent concentration; WB: 1:10000 - 1:50000; IP: 1:30; FC: 1:10 - 1:100; IHC-P: 1:250 - 1:500; ICC/IF: 1:250 - 1:500
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Does not react with Mouse, Rat
|Homo sapiens major histocompatibility complex, class I, A (HLA-A), transcript variant 1 (A*03:01:0:01 allele)|
|HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the alpha1 and alpha2 domains, which both bind the peptide, exon 4 encodes the alpha3 domain, exon 5 encodes the transmembrane region, and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described. [provided by RefSeq]. |
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Western blot - Anti-HLA A [EP1395Y] antibody; Anti-HLA A [EP1395Y] antibody at 1/10000 dilution + Raji cell lysate at 10 µg.Secondary.Goat anti rabbit IgG HRP conjugated at 1/2000 dilution.Predicted band size : 41 kDa.Observed band size : 41 kDa.
Anti-HLA A [EP1395Y] antibody; Ab52922 at 1/250 dilution staining human tonsil; paraffin embedded.
Immunocytochemistry/ Immunofluorescence - Anti-HLA A [EP1395Y] antibody; ICC/IF image of ab52922 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry - Anti-HLA A [EP1395Y] antibody; Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.