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CD34 Antibody (MEC 14.7)

Catalog # NB600-1071 | Novus Biologicals a Bio-Techne Brand
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NB600-1071
NB600-1071SS

Key Product Details

Species Reactivity

Mouse, Human (Negative)

Applications

ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # MEC 14.7

Concentration

1 mg/ml

Product Summary for CD34 Antibody (MEC 14.7)

Immunogen

Murine transformed endothelioma cell line t-end.

Reactivity Notes

This antibody does not detect Human CD34 and is an excellent tool for marking host derived endothelial cells/vasculature in human cancer xenografts on mouse.

Localization

Membrane; Single-pass type I membrane protein.

Marker

Hematopoietic Stem Cell Marker

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for CD34 Antibody (MEC 14.7)

Immunocytochemistry/Immunofluorescence: CD34 Antibody (MEC 14.7) [NB600-1071] - CD34 antibody was tested in WEHI-3 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Immunohistochemistry-Paraffin: CD34 Antibody (MEC 14.7) [NB600-1071] - Analysis of a FFPE tissue section of mouse small intestine using rat anti-mouse CD34 (clone MEC 14.7) at 1:100 dilution. The signal was developed using HRP-conjugated anti-rat secondary with DAB reagent which followed counterstaining of nuclei using hematoxylin. This antibody specifically labelled the endothelial cells in blood vessels located primarily in the sub-mucosa, and of that of the mucosa muscularis and the mucosal lacteal.
Flow Cytometry: CD34 Antibody (MEC 14.7) [NB600-1071] - CD34 (MEC 14.7) antibody was tested at 1:250 in WEHI-3 cells with DyLight 488 (green) alongside a matched isotype control (black).

Applications for CD34 Antibody (MEC 14.7)

Application
Recommended Usage

ELISA

1:100-1:2000

Flow Cytometry

1 ug per million cells

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Immunohistochemistry

1:100

Immunohistochemistry-Frozen

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1:10-1:500

Western Blot

1:250
Application Notes
This CD34 (MEC 14.7) antibody is useful for Immunohistochemistry (on both paraffin-embedded and frozen sections), Flow Cytometry, Immunocytochemistry/Immunofluorescence, Western blot, Immunoprecipitation and ELISA. Antigen retrieval is required for IHC-Paraffin.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 26 publications using NB600-1071 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD34

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

Alternate Names

CD34, HPCA1

Gene Symbol

CD34

Product Documents for CD34 Antibody (MEC 14.7)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for CD34 Antibody (MEC 14.7)

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.

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Protocols

View specific protocols for CD34 Antibody (MEC 14.7) (NB600-1071):

[[URL:https://www.novusbio.com/products/cd34-antibody-mec-147_nb600-1071]][[C… Antibody]]
Immunohistochemistry-paraffin embedded sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer pH 6.0 then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench top for 30 minutes.

Staining:
1. Wash sections in dH2O three times for 5 minutes each.
2. Wash section in wash buffer (1X PBS/0.1% Tween-20 (1X PBST)) for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1X PBST, 5% goat serum) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul primary antibody diluted in 1X PBST, 5% goat serum to each section. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated secondary antibody, diluted in 1X PBST, 5% goat serum. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Striptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in dH2O.
12. Counterstain sections in hematoxylin.
13. Wash sections in dH2O two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
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