CD34 Overexpression Lysate

Western Blot: CD34 Overexpression Lysate (Adult Normal) [NBP2-08650] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD34.

• Reactivity: Hu
• Applications: WB

CD34 Overexpression Lysate Summary

• Description:

  CD34 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204446
  
  Accession#: NM_001025109
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: CD34

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 40.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for CD34 Overexpression Lysate

• 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. Lysates are guaranteed for 6 months from date of receipt.

FAQs for CD34 Lysate (NBP2-08650). (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