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- Ab00127-1.1 Anti-CD34 [My10]
- Human
- Mouse IgG1
- Purified
- In Stock
- Ab00127-23.0 Anti-CD34 [My10]
- Human
- Rabbit IgG
- Purified
- In Stock
Recombinant monoclonal antibody to CD34. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma My10.
UniProt Accession Number of Target Protein: P28906
Alternative Name(s) of Target: Hematopoietic progenitor Ag; Hematopoietic progenitor cell antigen CD34; CD34
Immunogen: KG-1a Human myeloblastic leukemic cell line (immature human myeloid cell line) from a patient with nonlymphocytic leukemia.
Specificity: Human CD34, an ~105 - 120 kDa type 1 transmembrane glycoprotein expressed on the surface of most human hematopoietic stem and progenitor cells (HSPCs) as well as on mesenchymal stem cells, embryonic fibroblasts, endothelial cells, neurons, and some tumor cell lines.
Antibody first published in:
Civin CI, Strauss LC, Brovall C, Fackler MJ, Schwartz JF, Shaper JH. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984 Jul;133(1):157-65. PMID:6586833
Note on publication:
Describes the making of the antibody, shows it recognised ~115kDa My10 protein using immunoprecipitation and flow cytometry.
Flow-cytometry using anti-CD34 antibody My10 (Ab00127) Human leukocytes were stained with an isotype control (Ab00102-23.0, panel A) or the rabbit-chimeric version of My10 (Ab00127-23.0, 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 (ab150073) and cells analysed on a FACSCanto flow-cytometer.
Immunohistochemical staining of human placenta tissue using anti-CD34 antibody (Ab00127) My10 Anti-CD34 staining of paraffin embedded human placenta tissue using the rabbit-chimeric version of My10 (Ab00127-23.0). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer (Dako, cat. #X0909). Primary antibody incubation with Ab00127-23.0 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody (Dako cat#K4009) 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 (Ab00127) Jurkat cell extract (35µg protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of My10 (Ab00127-23.0) 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. Ab00127-23.0 successfully detected both isoforms of human CD34.