Recombinant Anti-c-Kit antibody [EPR25707-134] (ab283653)
![Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-477108-anti-c-kit-antibody-epr25707-134-western-blot-human.jpg "Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25707-134] to c-Kit
- Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-c-Kit antibody [EPR25707-134]
See all c-Kit primary antibodies -
Description
Rabbit monoclonal [EPR25707-134] to c-Kit -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEL, HL-60, Jurkat and Saos-2 whole cell lysates. IHC-P: Human colon, Human hyperplastic prostate, Human colon cancer and Human breast cancer tissues. ICC/IF: HEL cells. Flow Cyt: HL-60 and HEL cells. IP: Saos-2 whole cell lysate.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR25707-134 -
Isotype
IgG -
Research areas
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283653 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 |
|---|---|---|
| WB |
1/1000. Predicted molecular weight: 109 kDa.
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| IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
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| Flow Cyt |
1/500.
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| IP |
1/30.
|
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| ICC/IF |
1/250.
|
| Notes |
|---|
|
WB
1/1000. Predicted molecular weight: 109 kDa. |
|
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
Flow Cyt
1/500. |
|
IP
1/30. |
|
ICC/IF
1/250. |
Target
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Function
Tyrosine-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 specificity
Isoform 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 disease
Piebald trait
Gastrointestinal stromal tumor
Testicular germ cell tumor
Leukemia, acute myelogenous -
Sequence similarities
Belongs 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
modificationsUbiquitinated 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 localization
Cell membrane and Cytoplasm. Detected in the cytoplasm of spermatozoa, especially in the equatorial and subacrosomal region of the sperm head. - Information by UniProt
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Database links
- Entrez Gene: 3815 Human
- Omim: 164920 Human
- SwissProt: P10721 Human
- Unigene: 479754 Human
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Alternative names
- C Kit antibody
- c-Kit antibody
- c-Kit Ligand antibody
see all
Images
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Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)All lanes : Anti-c-Kit antibody [EPR25707-134] (ab283653) at 1/1000 dilution
Lane 1 : HEL (human erythroleukemia erythroblast) whole cell lysate
Lane 2 : HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 4 : Saos-2 (human osteosarcoma epithelial) whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 109 kDa
Observed band size: 120,140-160 kDa why is the actual band size different from the predicted?
Exposure time: 125 secondsBlocking buffer: 5% NFDM/TBST.
Low expression: HL-60 (PMID: 29127384) and Jurkat (PMID: 22140461).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-1-anti-c-kit-antibody-epr25707-134-immunhistochemistry-human-colon.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling c-Kit with ab283653 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining in lamina propria cells of human colon (PMID: 23276179). The section was incubated with ab283653 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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![Immunocytochemistry/ Immunofluorescence - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-5-anti-c-kit-antibody-epr25707-134-immuncytochemistry-hel-cells.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-c-Kit antibody [EPR25707-134] (ab283653)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEL cells labelling c-Kit with ab283653 at 1/250 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing membranous staining in HEL cells. Negative control: HL-60 (PMID: 29127384) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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![Flow Cytometry - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-6-anti-c-kit-antibody-epr25707-134-flow-cytometry-hl-60.jpg)
Flow Cytometry - Anti-c-Kit antibody [EPR25707-134] (ab283653)Flow cytometric analysis of HL-60 (Human Acute Promyelocytic Leukemia promyeloblast, Left)/ Saos-2 (human osteosarcoma epithelial, Right) cells labelling c-Kit with ab283653 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 was used as the secondary antibody. Negative control: HL-60 (PMID: 29127384). Gated on viable cells.
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![Immunoprecipitation - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-8-anti-c-kit-antibody-epr25707-134-immunprecipitation-soas-2-human.jpg)
Immunoprecipitation - Anti-c-Kit antibody [EPR25707-134] (ab283653)c-Kit was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial) whole cell lysate 10 μg with ab283653 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab283653 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Saos-2 (human osteosarcoma epithelial) whole cell lysate 10 μg
Lane 2: Saos-2 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283653 in Saos-2 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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![Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-477110-anti-c-kit-antibody-epr25707-134-hel-human.jpg)
Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)All lanes : Anti-c-Kit antibody [EPR25707-134] (ab283653) at 1/1000 dilution
Lane 1 : HEL (human erythroleukemia erythroblast) whole cell lysate
Lane 2 : HEL whole cell lysate treated with PNGase F
Lane 3 : Saos-2 (human osteosarcoma epithelial) whole cell lysate
Lane 4 : Saos-2 whole cell lysate treated with PNGase F
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 109 kDa
Observed band size: 120,140-160 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking buffer: 5% NFDM/TBST.
The difference in target band size observed in HEL and Saos-2 cell lysates is caused by glycosylation.
This blot was developed using a higher sensitivity ECL substrate.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-2-anti-c-kit-antibody-epr25707-134-immunhistochemistry-human-hyperplastic-prostate.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)Immunohistochemical analysis of paraffin-embedded Human hyperplastic prostate tissue labelling c-Kit with ab283653 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on stromal cells of human hyperplastic prostate (PMID: 23276179). The section was incubated with ab283653 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
![Flow Cytometry - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-7-anti-c-kit-antibody-epr25707-134-flow-cytometry-hel-human.jpg)
Flow Cytometry - Anti-c-Kit antibody [EPR25707-134] (ab283653)Flow cytometric analysis of HEL (human erythroleukemia erythroblast) cells labelling c-Kit with ab283653 (Red) at 1/500 dilution compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody. Gated on viable cells.
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![Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-477113-anti-c-kit-antibody-epr25707-134-hel-saos-2-jurkat-human.jpg)
Western blot - Anti-c-Kit antibody [EPR25707-134] (ab283653)All lanes : Anti-c-Kit antibody [EPR25707-134] (ab283653) at 1/1000 dilution
Lane 1 : HEL (human erythroleukemia erythroblast) whole cell lysate
Lane 2 : Saos-2 (human osteosarcoma epithelial) whole cell lysate
Lane 3 : HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate
Lane 4 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 109 kDa
Observed band size: 120,140-160 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking buffer: 5% NFDM/TBST.
Low expression: HL-60 (PMID: 29127384) and Jurkat (PMID: 22140461).
Bis-tris gel was used in the blot.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-3-anti-c-kit-antibody-epr25707-134-immunhistochemistry-human-colon-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labelling c-Kit with ab283653 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on stromal cells of human colon cancer (PMID: 23276179). The section was incubated with ab283653 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)](https://www.abcam.com/ps/products/283/ab283653/Images/ab283653-4-anti-c-kit-antibody-epr25707-134-immunhistochemistry-human-breast-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Kit antibody [EPR25707-134] (ab283653)Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling c-Kit with ab283653 at 1/100 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human breast cancer. The section was incubated with ab283653 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab283653 has not yet been referenced specifically in any publications.