Anti-c-Kit antibody [2B8] (Biotin) (ab25022)

Overview

  • Product nameAnti-c-Kit antibody [2B8] (Biotin)
    See all c-Kit primary antibodies
  • Description
    Rat monoclonal [2B8] to c-Kit (Biotin)
  • ConjugationBiotin
  • Tested applicationsSuitable for: IHC-Fr, Flow Cyt, Functional Studies, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    The details of the immunogen for this antibody are not available.

Applications

Our Abpromise guarantee covers the use of ab25022 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
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab18542-Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
Functional Studies Use at an assay dependent concentration. Can be used for cell homing studies, blockade of hematopoiesis and disruption fo melanocyte development.
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionTyrosine-protein kinase that acts as cell-surface receptor for the cytokine KITLG/SCF and plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. In response to KITLG/SCF binding, KIT can activate several signaling pathways. Phosphorylates PIK3R1, PLCG1, SH2B2/APS and CBL. Activates the AKT1 signaling pathway by phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase. Activated KIT also transmits signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. Promotes activation of STAT family members STAT1, STAT3, STAT5A and STAT5B. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KIT signaling is modulated by protein phosphatases, and by rapid internalization and degradation of the receptor. Activated KIT promotes phosphorylation of the protein phosphatases PTPN6/SHP-1 and PTPRU, and of the transcription factors STAT1, STAT3, STAT5A and STAT5B. Promotes phosphorylation of PIK3R1, CBL, CRK (isoform Crk-II), LYN, MAPK1/ERK2 and/or MAPK3/ERK1, PLCG1, SRC and SHC1.
  • Tissue specificityIsoform 1 and isoform 2 are detected in spermatogonia and Leydig cells. Isoform 3 is detected in round spermatids, elongating spermatids and spermatozoa (at protein level). Widely expressed. Detected in the hematopoietic system, the gastrointestinal system, in melanocytes and in germ cells.
  • Involvement in diseasePiebald trait
    Gastrointestinal stromal tumor
    Testicular germ cell tumor
    Leukemia, acute myelogenous
  • Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Ubiquitinated by SOCS6. KIT is rapidly ubiquitinated after autophosphorylation induced by KITLG/SCF binding, leading to internalization and degradation.
    Autophosphorylated on tyrosine residues. KITLG/SCF binding enhances autophosphorylation. Isoform 1 shows low levels of tyrosine phosphorylation in the absence of added KITLG/SCF (in vitro). Kinase activity is down-regulated by phosphorylation on serine residues by protein kinase C family members. Phosphorylation at Tyr-568 is required for interaction with PTPN11/SHP-2, CRK (isoform Crk-II) and members of the SRC tyrosine-protein kinase family. Phosphorylation at Tyr-570 is required for interaction with PTPN6/SHP-1. Phosphorylation at Tyr-703, Tyr-823 and Tyr-936 is important for interaction with GRB2. Phosphorylation at Tyr-721 is important for interaction with PIK3R1. Phosphorylation at Tyr-823 and Tyr-936 is important for interaction with GRB7.
  • Cellular localizationCell membrane and Cytoplasm. Detected in the cytoplasm of spermatozoa, especially in the equatorial and subacrosomal region of the sperm head.
  • Information by UniProt
  • Database links
  • Alternative names
    • C Kit antibody
    • c-Kit antibody
    • c-Kit Ligand antibody
    • CD117 antibody
    • Kit antibody
    • Kit Ligand antibody
    • KIT oncogene antibody
    • KIT proto oncogene receptor tyrosine kinase antibody
    • KIT_HUMAN antibody
    • Mast cell growth factor receptor antibody
    • Mast/stem cell growth factor receptor Kit antibody
    • MGF antibody
    • p145 c-kit antibody
    • PBT antibody
    • Piebald trait protein antibody
    • Proto oncogene c Kit antibody
    • Proto oncogene tyrosine protein kinase Kit antibody
    • Proto-oncogene c-Kit antibody
    • SCF Receptor antibody
    • SCFR antibody
    • soluble KIT variant 1 antibody
    • Steel Factor Receptor antibody
    • Stem cell factor receptor antibody
    • tyrosine protein kinase Kit antibody
    • Tyrosine-protein kinase Kit antibody
    • v kit Hardy Zuckerman 4 feline sarcoma viral oncogene homolog antibody
    • v kit Hardy Zuckerman 4 feline sarcoma viral oncogene like protein antibody
    • v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog antibody
    see all

Anti-c-Kit antibody [2B8] (Biotin) images

  • Immunostaining of mouse embryonic stem cells using c-Kit (Biotin) ab25022. The antibody bound to the cells was detected by treatment with Cy3-conjugated streptavidin (25 µg/ml). The image shows a fragment of an adherent colony of mouse embryonic stem cell-derived hepatocyte precursor cells stained with the ab25022 antibody and subsequently treated with Cy3-conjugated streptavidin. Cell nuclei were stained with DAPI. The Cy3 fluorescence pattern suggests the predominant expression of c-Kit in actively proliferating cells in the middle of colonies. Please see abreview for ICC/IF protocol details.

    See Abreview

  • ab25022 staining c-Kit (red) in murine testis tissue sections by Immunohistochemistry (Frozen sections).

    Tissue was fixed with paraformaldehyde and blocked with 2% BSA for 20 minutes at 20°C. Samples were incubated with primary antibody (1/100 in 2% BSA + 0.005% Triton X-100 in PBS) for 16 hours at 4°C. Avidin-FITC was used for detection.

    Nuclei were stained with DAPI (blue).

    See Abreview

  • Flow Cytometry analysis of C57BL/6 mouse bone marrow cells labeling c-Kit with ab25022 at 0.3 μg/106 cells dilution. Rat Anti-Mouse CD45R-PE followed by Streptavidin-FITC was used as the secondary antibody. 

References for Anti-c-Kit antibody [2B8] (Biotin) (ab25022)

This product has been referenced in:
  • Drummond CA  et al. Reduction of Na/K-ATPase affects cardiac remodeling and increases c-kit cell abundance in partial nephrectomized mice. Am J Physiol Heart Circ Physiol 306:H1631-43 (2014). Read more (PubMed: 24748592) »
  • Drobinskaya I  et al. Scalable selection of hepatocyte- and hepatocyte precursor-like cells from culture of differentiating transgenically modified murine embryonic stem cells. Stem Cells 26:2245-56 (2008). ICC/IF ; Mouse . Read more (PubMed: 18556507) »

See all 2 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Sample Mouse Tissue sections (Left Ventricle)
Specification Left Ventricle
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 20 2014


Earlier we spoke of the non-specific staining and lack of expected, positive staining in your IHC sections. We discussed attempting antigen retrieval at 95-100C rather than 80C and trying some alternative buffers (Ex. Tris-EDTA) and potentially an...

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Thank you for contacting us.

I am sorry to hear thatone of the antibodies is not working as expected.

According to the image you had sent, it seems that the c-kit antibody (ab25022) is not working correctly. However the vimentin ant...

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Thank you for contacting us.

Although we have not determined the affinity of this antibody against the rat orthologue, we are pleased to let you know that we guarantee this c-Kit antibody ab25022 to work with rat samples. We are happy to rea...

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Thank you for your enquiry. I have looked through our stock of c-Kit antibodies and unfortunately have not found one where references we know of support its use to block receptor function. I am sorry I could not be of more help in this instance. Please...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Rat Cell (Cardiac progenitor cell)
Specification Cardiac progenitor cell
Preparation Cell harvesting/tissue preparation method: Cardiomycyte isolation
Sample buffer: Kreb's
Fixation PBS
Permeabilization No
Gating Strategy Cardiac Progenitor Cells
Username

Abcam user community

Verified customer

Submitted Oct 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (testis)
Specification testis
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Feb 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Embryonic stem cell-derived hepatocyte precursors)
Specification Embryonic stem cell-derived hepatocyte precursors
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Irina Drobinskaya

Verified customer

Submitted Feb 17 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"