Overview

  • Product name

    Anti-c-Kit (phospho Y703) antibody
    See all c-Kit primary antibodies
  • Description

    Rabbit polyclonal to c-Kit (phospho Y703)
  • Host species

    Rabbit
  • Specificity

    ab62154 detects endogenous levels of c-Kit only when phosphorylated at tyrosine 703 (Human: Tyr703; Mouse: Tyr702).
  • Tested applications

    Suitable for: WB, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic phosphopeptide derived from human c-Kit around the phosphorylation site of tyrosine 703 (A-L-YP-K-N).

  • Positive control

    • Human breast carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab62154 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 Use at an assay dependent concentration.
IHC-P 1/50 - 1/100.
ELISA 1/5000.

Target

  • 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
    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 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
  • 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

Images

  • Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue using c-Kit (phospho Y703) antibody (ab62154) at 1/50 - 1/100 dilution, in the presence (right panel) and absence (left panel) of immunising phosphopeptide.
  • All lanes : Anti-c-Kit (phospho Y703) antibody (ab62154)

    Lane 1 : Colon cancer lysate (human)
    Lane 2 : U2OS lysate (human)
    Lane 3 : Renal cancer cell line (human)


    Secondary
    All lanes : Swine ati-rabbit HRP at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 6 minutes

    See Abreview

References

This product has been referenced in:

  • Zhao X  et al. Lactobacillus plantarum YS2 (yak yogurt Lactobacillus) exhibited an activity to attenuate activated carbon-induced constipation in male Kunming mice. J Dairy Sci 102:26-36 (2019). Read more (PubMed: 30527985) »
  • Yang H  et al. Predictive and prognostic value of phosphorylated c-KIT and PDGFRA in advanced non-small cell lung cancer harboring ALK fusion. Oncol Lett 17:3071-3076 (2019). Read more (PubMed: 30867736) »
See all 6 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

It is my pleasure to help.

Please let me know if you have any further questions or if there are any ways in which Abcam might help you in the future.

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Answer

This will not be a problem. Our accounts team can gladly arrange thisfor you. Please have your purchase department contact our accounts team at the email address which was included in the credit note details or by telephone at 888-772-2226.



I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question

  Below is my respond to your questions.   As you had ordered a single vial in July followed by 4 more in August, am I correct in assuming that the first vial did give good specific result but subsequent ones did not? Did you test each the vials from you August order to determine whether this was vial specific problem? I ask as these facts are important for us to determine if there was something which may have happened to the subsequent vials which might have affected them. The first vial was tested on breast carcinoma tissue.  There was some non specific staining in nucleus of cancer tissue as well as normal tissue.  However, we decided to go ahead with our sample (Adenoid cystic carcinoma of Salivary gland) WITH the isotype control staining.  The isotype control should help us identify the non-specific staining and we can read the signal from specific antibody.  We ordered one vial of isotype control and 4 vials of p-c-kit antibody, the same lot number with the first one.  We pooled the entire antibody and stain all 35 slides.  The isotype control was used in the same concentration and uses the same protocol for 6 slides.   The test sample and isotype control were positive in cancer tissue, normal tissue, nucleus, cytoplasm and cell membrane for all slides with stronger signals than the test samples.  The pathologist was not able to interpret the result from this staining.    Thank you for providing information regarding the dilution, tissue, control and detection system which you have used with this product. If you don't mind, would you be able to tell me the antigen retrieval, blocking, antibody incubation and washing conditions which you have used as well. These details will help us should we need to have the product re-submitted for quality control. I am attached the protocol that UCSF Cancer Center IHC core use for my staining with this mail.   I would also like to remind you that if any Abcam product does not work for you, please contact us immediately. Our products are covered by our six month Abpromise in which we not only provide support but are able to replace or refund in that period if they are being used in applications and species listed on the datasheet.  We ordered one vial in 7/23/12.  We tested on breast carcinoma tissue with 1:100, 1:50 and 1:25 on 8/2/12.  The pathologist saw only 1+ signal from the sample.  She asked us to perform again with higher concentration to increase the signal.  We performed the IHC again with 1:20, 1:10 and 1:5 on 8/8/12.  This time, she saw the 2+to 3+ signals in 1:10 dilution. I ordered the antibody again, along with isotype control on 8/13/12.  We received the antibody on 8/21/12.  The slides were stained on 8/23/12 and reviewed by pathologist on 8/27/12.  I reported the problem to you the same day-yesterday.     We do understand the financial pressures felt by researchers these days which is why we are happy to have Abpromise in place. While we are able to refund Abcam products, under our terms and conditions we cannot compensate for additional costs. " Any claim relating to products shall be limited to replacement or refund of original purchase price paid. " I do look forward to hearing from you these additional protocol details. If you could please confirm the Abcam order confirmation numbers or POs, this would be of great help in moving forward as well. The first order PO is B000094576 received on 7/23/12. The second order PO is B000104345 received on 8/21/12.   Thank you in advance for your help. Please let me know if you have any other questions.  

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Answer

Thank you for contacting us. Your credit note ID are xxxx and xxxxxx. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question

I have a complain about your product phospho c-kit antibody (ab62154).  I have been working with this antibody, using the different dilution (1:5 to 1:100) and different breast carcinoma tissue as well as different DAP developing time.  We have the best dilution and last week, along with isotype control staining, we completed the staining for my samples.  Unfortunately, we cannot use any of our data from the staining due to several reasons.   1.  The antibody are stain non-specific.  The positive signal should be only on the cell membrane, however they are stain all in nucleus, cytoplasm and cell membrane.  The pathologist is not able to evaluate the significant of this staining. 2.  The isotype staining is very strong.  The signal from isotype staining is much more than the specific antibody.  This make it impossible for interpret any signal from specific antibody.   I have invested a lot of my time, money and energy for this staining.  I spent the money for 6 tubes of antibodies, over $1200 for IHC core to stain the sample for optimization and test, $300 for section preparation and most of all, my FFPE tissue sample which is the rare cases.  We have only 33 cases for the last 20 years archive.    I am writing this e-mail to ask for the reimbursement of the money that I spent for at least the antibody part from Abcam.  Would you please consider refund the money for me?  As you know, all the lab has very hard time with money issue now.  Spending more than $6000 and time is not acceptable for me.  I can send in the slide if you would like to, there is no use in the lab.

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Answer

My name is Keith. John is away for a few weeks and as such I will be helping you with this problem.

First, I want to express my sympathy to you. I do understand the frustration which comes from this type of problem and want to help resolve this issue.

As you had ordered a single vial in July followed by 4 more in August, am I correct in assuming that the first vial did give good specific result but subsequent ones did not? Did you test each the vials from you August order to determine whether this was vial specific problem? I ask as these facts are important for us to determine if there was something which may have happened to the subsequent vials which might have affected them.

Thank you for providing information regarding the dilution, tissue, control and detection system which you have used with this product. If you don't mind, would you be able to tell me the antigen retrieval, blocking, antibody incubation and washing conditions which you have used as well. These details will help us should we need to have the product re-submitted for quality control.

I would also like to remind you that if any Abcam product does not work for you, please contact us immediately. Our products are covered by our six month Abpromise in which we not only provide support but are able to replace or refund in that period if they are being used in applications and species listed on the datasheet.

We do understand the financial pressures felt by researchers these days which is why we are happy to have Abpromise in place. While we are able to refund Abcam products, under our terms and conditions we cannot compensate for additional costs. "Any claim relating to products shall be limited to replacement or refund of original purchase price paid." (https://www.abcam.com/index.html?pageconfig=terms)

I do look forward to hearing from you these additional protocol details. If you could please confirm the Abcam order confirmation numbers or POs, this would be of great help in moving forward as well. Please let me know if you have any

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Answer

Thank you for your reply.

We do have a few other rabbit polyclonal isotype controls that are at a higher concentration, these are:

https://www.abcam.com/Rabbit-polyclonal-IgG-ab27478.html
https://www.abcam.com/Rabbit-IgG-ChIP-Grade-ab37415.html



If there is anything else I can help you with, or if there are any other products I can help you select, please let me know.

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Answer

Thank you for your reply.

As this is a rabbit polyclonal antibody, the exact clonality of the antibody cannot be determined. However we do have isotype controls that would work for you, please see the link below:


https://www.abcam.com/Rabbit-polyclonal-IgG-ab27472.html



If there is anything else I can help you with, please let me know.

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Answer

Thank you for contacting Abcam.

I have attached the IHC-P protocol that was used to test ab62154 and currently we do not know if any publications that have used this antibody in IHC-P, that does not mean there are not any, just that we do not know.

Also, I do not have any further information about the breast cancer samples that was used.

I am sorry that I could not help further in this matter, but please let me know if you have any further questions.

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Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cancer (tissue lysate), U2OS (cell lysate) a)
Loading amount
20 µg
Specification
Colon cancer (tissue lysate), U2OS (cell lysate) a
Gel Running Conditions
Reduced Denaturing (Pierce precise protein gel 4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Mr. Antibody Solutions

Verified customer

Submitted Apr 20 2011

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