CD34 Antibody (My10) - Chimeric - Azide and BSA Free

Western Blot: CD34 Antibody (My10) - Chimeric [NBP2-62554] - Western Blot using anti-CD34 antibody My10. Jurkat cell extract (35ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of My10 (NBP2-62554) at 0.1 ug/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. NBP2-62554 successfully detected both isoforms of human CD34.

Immunohistochemistry: CD34 Antibody (My10) - Chimeric [NBP2-62554] - 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 (NBP2-62554). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation with NBP2-62554 was carried out at 4 ug/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 ug/ml.

Flow Cytometry: CD34 Antibody (My10) - Chimeric [NBP2-62554] - Flow-cytometry using anti-CD34 antibody My10. Human leukocytes were stained with an isotype control (panel A) or the rabbit-chimeric version of My10 (NBP2-62554, panel B) at a concentration of 1 ug/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.

• Reactivity: Hu
• Applications: WB, Flow, IHC, IP
• Clone: My10
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

CD34 Antibody (My10) - Chimeric - Azide and BSA Free Summary

• Additional Information: Recombinant Monoclonal Antibody. Clone My10 was used by HLDA to establish CD designation.
• 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.
• Isotype: IgG Kappa
• Clonality: Monoclonal
• Host: Rabbit
• Gene: CD34
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:10 - 1:1000
  • Immunohistochemistry 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot
• Application Notes: This chimeric rabbit antibody was made using the variable domain sequences of the original murine IgG1 format, for improved compatibility with existing reagents, assays and techniques.

Packaging, Storage & Formulations

• Storage: Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.
• Buffer: PBS
• Preservative: 0.02% Proclin 300
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for CD34 Antibody (My10) - Chimeric - Azide and BSA Free

• CD34 antigenhematopoietic progenitor cell antigen CD34
• CD34 molecule
• CD34
• HPCA1

Background

CD34, also known as Cluster of Differentiation 34, is a transmembrane phosphoglycoprotein that is primarily known for being a surface marker for hematopoietic stem cells (HSCs) and progenitor cells (1,2). The CD34 protein consists of a heavily glycosylated extracellular domain, a single-pass transmembrane helix, and a cytoplasmic tail with PDZ binding motifs (1,3). Human CD34 has two protein isoforms consisting of 385 amino acids (aa) for the canonical isoform and 328 aa for isoform 2 (3,4). The canonical CD34 isoform has calculated theoretical molecular weight of 40 kDa and an observed molecular weight of ~115 kDa (1,3,4). L-Selectin (CD62L) is the most common ligand for CD34, but it has also been shown to bind CrkL (1,3).

CD34 has commonly been used as a marker for the diagnosis and classification of various diseases and pathologies including leukemia and solitary fibrous tumor (SFT) (2,5). In terms of immunohistochemistry and histopathology, CD34 has been the most common marker for SFT and is expressed in ~79% of cases (5). In addition to its use as a cell marker, CD34-postive (CD34+) hematopoietic stem cells have been used therapeutically in patients following radiation or chemotherapy due to their regenerative potential (6). There are several clinical trials showing promising results for CD34+ cell therapy for cardiovascular diseases including heart failure, ischemia, dilated cardiomyopathy, acute myocardial infarction, and angina (6). Besides hematopoietic lineages, CD34 is also expressed in non-hematopoietic cells including mesenchymal stem cells, endothelial cells and progenitors, fibrocytes, muscle satellite cells, and some cancer stem cells (1,3). While the clinical and cell therapy applications of CD34 as a cell marker is well documented, the function of CD34 is less understood but has been implicated in many cellular processes such as adhesion, proliferation, signal transduction, differentiation, and progenitor phenotype maintenance (1,3).

References

1. Sidney, L. E., Branch, M. J., Dunphy, S. E., Dua, H. S., & Hopkinson, A. (2014). Concise review: evidence for CD34 as a common marker for diverse progenitors. Stem cells (Dayton, Ohio), 32(6), 1380-1389. https://doi.org/10.1002/stem.1661

2. Krause, D. S., Fackler, M. J., Civin, C. I., & May, W. S. (1996). CD34: structure, biology, and clinical utility. Blood, 87(1), 1-13

3. Kapoor, S., Shenoy, S. P., & Bose, B. (2020). CD34 cells in somatic, regenerative and cancer stem cells: Developmental biology, cell therapy, and omics big data perspective. Journal of cellular biochemistry, 121(5-6), 3058-3069. https://doi.org/10.1002/jcb.29571

4. Uniprot (P28906)

5. Davanzo, B., Emerson, R. E., Lisy, M., Koniaris, L. G., & Kays, J. K. (2018). Solitary fibrous tumor. Translational gastroenterology and hepatology, 3, 94. https://doi.org/10.21037/tgh.2018.11.02

6. Prasad, M., Corban, M. T., Henry, T. D., Dietz, A. B., Lerman, L. O., & Lerman, A. (2020). Promise of autologous CD34+ stem/progenitor cell therapy for treatment of cardiovascular disease. Cardiovascular research, 116(8), 1424-1433. https://doi.org/10.1093/cvr/cvaa027

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

FAQs for CD34 Antibody (NBP2-62554). (Showing 1 - 1 of 1 FAQ).

1. I wonder if you have a CD105 or CD34 antibody suitable for IHC that is specific for human and do not bind mouse?
   • We do not have any anti-human CD34 or CD105 antibodies that are confirmed to NOT detect the mouse protein. When we have tested an antibody and confirmed that it will not react with mouse samples, we will add Mu(-) to the datasheet, and unfortunately all of our CD105 and CD34 antibodies will either detect the mouse protein, or they have not been used in mouse samples before.

Bioinformatics

• Gene Symbol: CD34