Anti-CD34 [My10]

• Catalog Number: orb411545
• Category: Antibodies
• Description: Mouse monoclonal antibody to CD34
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: My10
• Tested applications: FC, IHC, IP
• Reactivity: Human
• Isotype: IgG1
• Immunogen: KG-1a Human myeloblastic leukemic cell line (immature human myeloid cell line) from a patient with nonlymphocytic leukemia.
• Concentration: 1 mg/ml
• Purity: Purified
• Conjugation: Unconjugated
• Target: CD34
• UniProt ID: P28906
• 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: PBS with 0.02% Proclin 300.
• Alternative names: Hematopoietic progenitor Ag; Hematopoietic progeni
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow-cytometry using anti-CD34 antibody My10 Human leukocytes were stained with an isotype control (orb256390, panel A) or the rabbit-chimeric version of My10 (orb411546, panel B) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody and cells analysed on a FACSCanto flow-cytometer.

Immunohistochemical staining of human placenta tissue using anti-CD34 antibody My10 Anti-CD34 staining of paraffin embedded human placenta tissue using the rabbit-chimeric version of My10 (orb411546). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation with orb411546 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody for 20 mins followed by DAB (3, 3′-diaminobenzidine), and counter-staining with haemotoxylin. Strong endothelial staining of within villi may be observed. Recommended concentration, 2-4 µg/ml.

Western Blot using anti-CD34 antibody My10 Jurkat cell extract (35 µg protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of My10 (orb411546) at 0.1 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for the canonical isoform of CD34 is 40.9kDa, though a second isoform at 35.2kDa is also present, with a 56 amino acid truncation at the C-terminus of the protein (Uniprot ID: Q64314). CD34 is also glycosylated at several positions. orb411546 successfully detected both isoforms of human CD34.