Anti-Hu CD45 Purified

• Catalog Number: orb44725
• Category: Antibodies
• Description: Mouse monoclonal antibody to CD45
• Clonality: Monoclonal
• Clone Number: HI30
• Tested applications: FC, ICC, IHC-Fr, IHC-P, IP, WB
• Reactivity: Human, Primate
• Isotype: Mouse IgG1 kappa
• Immunogen: human peripheral blood leukocytes
• Antibody Type: Primary Antibody
• Concentration: 1 mg/ml
• Dilution range: Western blotting: Recommended dilution: 1 μg/ml; non-reducing conditions, band around 200-250 kDa; positive control: JURKAT human leukemia T-cell lysate, Kg-1a human leukemia cell lysate; non-reducing conditions, 6% separating gel. Flow cytometry: Recommended dilution: 1-4 µg/ml
• Purity: Purified by protein-G affinity chromatography.
• Conjugation: Unconjugated
• Target: CD45
• Entrez: 5788
• UniProt ID: P08575
• RRID: AB_10999116
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
• Alternative names: anti CD 45 antibody, anti CD45 antibody, anti CD45
• Note: For research use only
• Application notes: Western blotting: non-reducing conditions; recommended dilution: 1 μg/ml; 6-7% gel recommended (band around 180-250 kDa).Flow cytometry: Recommended dilution: 1-4 µg/ml
• Expiration Date: 12 months from date of receipt.

Flow cytometry analysis (surface staining) of human peripheral blood cells with anti-human CD45 (HI30) purified, GAM-APC.

Anti-Hu CD45 Purified (clone HI30) works in WB application under non-reducing conditions. Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of Jurkat, Raji, HeLa, and HEK293T cell lines, mixed and heated (100°C, 5 min) with reducing and non-reducing SDS-loading buffer. Samples were resolved using 7% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed with mouse IgG1 monoclonal antibody HI30 (1 µg/ml), followed by IRDye 800CW Goat-anti-Mouse IgG (green). Multiplex fluorescent Western blot detection was performed. CD45 molecules were detected at ~180-250 kDa in Jurkat and Raji cell lines.