Overview

  • Product name
    Anti-CD34 antibody [MEC 14.7]
    See all CD34 primary antibodies
  • Description
    Rat monoclonal [MEC 14.7] to CD34
  • Specificity
    Recognizes mouse CD34.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IHC-Fr, IP, IHC-P, WB, IHC-FrFl, ELISAmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to Mouse CD34. Specifically, the murine endothelioma cell line tEnd.

  • Positive control
    • Mouse lung tissue.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab8158 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
Flow Cyt 1/50.

See Sho et al and Garlanda et al.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-Fr 1/50.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 0.5 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Staining with this method can be difficult, it has been reported to us that milder fixation methods for paraffin sections like using zinc solution work well (unfortunately we have no further detailed instructions of this fixation method).

WB Use a concentration of 5 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 41 kDa).
IHC-FrFl Use at an assay dependent concentration. PubMed: 20382702
ELISA Use at an assay dependent concentration.

Target

  • Function
    Possible adhesion molecule with a role in early hematopoiesis by mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Could act as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components. Presents carbohydrate ligands to selectins.
  • Tissue specificity
    Selectively expressed on hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues.
  • Sequence similarities
    Belongs to the CD34 family.
  • Developmental stage
    On early hematopoietic progenitor cells.
  • Post-translational
    modifications
    Highly glycosylated.
    Phosphorylated on serine residues by PKC.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD34 antibody
    • CD34 antigen antibody
    • CD34 molecule antibody
    • CD34_HUMAN antibody
    • Cluster designation 34 antibody
    • Hematopoietic progenitor cell antigen CD34 antibody
    • HPCA1 antibody
    • Mucosialin antibody
    • OTTHUMP00000034733 antibody
    • OTTHUMP00000034734 antibody
    see all

Images

  • Anti-CD34 antibody [MEC 14.7] (ab8158) at 1 µg/ml + Mouse Lung Membrane Tissue Lysate (ab171830) at 50 µg

    Secondary
    Goat Anti-Rat IgG H&L (HRP) (ab97057) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size : 41 kDa
    Observed band size : 80 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    The main band is as expected at 80 kDa since the target is heavily glycosylated and phosphorylated.

  • Immunohistochemical analysis of murine lung tissue, staining CD34 with ab8158.

    Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in diluent) for 16 hours at 4°C. An undiluted HRP-conjugated horse anti-rat polyclonal IgG was used as the secondary antibody.

    See Abreview

  • ab8158 staining CD34 in mouse lung endothelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 1% BSA for 30 minutes at 4°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An undiluted Alexa Fluor® 555-conjugated goat anti-rat IgG polyclonal was used as the secondary antibody.

    See Abreview

  • IHC image of CD34 staining in Mouse brain FFPE section, performed on a BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8158, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
  • ab8158 staining CD34 in mouse mammary gland tissue sections by Immunohistochemistry (IHC-P - paraffin-embedded sections). Tissue was fixed with methacarnoy and blocked with 4% BSA for 60 minutes; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100) for 16 hours. A undiluted HRP-conjugated goat anti-rat IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab8158 staining mouse brain tissue sections by IHC-P.  Tissue sections were paraformaldehyde fixed and blocked with 0.5% Perkin-Elmer TNB Blocking Buffer for 30 minutes at 25°C.  The primary antibody was diluted 1/100 and incubated for 18 hours at 4°C.  An biotin conjugated goat anti-rat was used as the secondary

    Photomicrograph demonstrates CD34 in red and collagen type IV (ab19808) in blue in normal, adult brain vessels. Tissue was perfusion-fixed and cut into 15µm slide-mounted cryostat sections (i.e., lightly fixed, but not paraffin embedded).

    See Abreview

  • Ab8158 at a dilution of 1/50 staining CD34 from mouse lung cells by Immunocytochemistry. The antibody was incubated with the sample for 1 hour and then detected using an Alexa-Fluor 488® goat anti-rat antibody. The staining for CD34 is shown in green. The samples were also stained for Smooth Muscle Actin (red stain).

    This image is courtesy of an Abreview submitted on 8 February 2006.

    See Abreview

References

This product has been referenced in:
  • Fu W  et al. Differential effects of recombinant human endostatin treatment on differentiated and undifferentiated blood vessels in Lewis lung cancer. Oncol Lett 13:196-200 (2017). Read more (PubMed: 28123541) »
  • Komla-Ebri D  et al. Tyrosine kinase inhibitor NVP-BGJ398 functionally improves FGFR3-related dwarfism in mouse model. J Clin Invest 126:1871-84 (2016). Read more (PubMed: 27064282) »

See all 131 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (sarcoma tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Permeabilization
No
Specification
sarcoma tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 21°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

Submitted Aug 24 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (melanoma cell line xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Permeabilization
No
Specification
melanoma cell line xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 16 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Cytospined Bone marrow cells)
Permeabilization
No
Specification
Cytospined Bone marrow cells
Fixative
Methanol
Username

Dr. Francois Daubeuf

Verified customer

Submitted Apr 27 2015

Application
Flow Cytometry
Fixation
None, cells were used fresh
Permeabilization
No
Sample
Mouse Cell (Cells from bone marrow (C57b6 mice))
Specification
Cells from bone marrow (C57b6 mice)
Gating Strategy
Alive cells (DAPI-), blood cells exluded
Preparation
Cell harvesting/tissue preparation method: Bone marrow from Femur was extracted with PBS-EDTA 3mM, 2 hours before flow cytometry analysis
Sample buffer: PBS-EDTA 3mM
Username

Dr. Francois Daubeuf

Verified customer

Submitted Feb 24 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (lung)
Specification
lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Nov 10 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate (pH 6)
Sample
Mouse Tissue sections (Metastatic Lung)
Specification
Metastatic Lung
Permeabilization
No
Fixative
Formaldehyde
Username

Dr. Victoria Thompson

Verified customer

Submitted Feb 05 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 4°C
Sample
Mouse Cell (Lung Endothelial)
Specification
Lung Endothelial
Permeabilization
No
Fixative
Formaldehyde
Username

Dr. Victoria Thompson

Verified customer

Submitted Feb 05 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.1M Citrate buffer pH 6.5
Sample
Rat Tissue sections (rat pancreas)
Specification
rat pancreas
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Specification
Liver
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
Yes - Tween-20 1%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 16 2013



J’ai contacté le laboratoire qui m’a confirmé que tous nos lots d’ab8158 sont testés en immunohistochimie sur coupes paraffines.

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