Overview

  • Product name

    Anti-Src (phospho S75) antibody
    See all Src primary antibodies
  • Description

    Rabbit polyclonal to Src (phospho S75)
  • Host species

    Rabbit
  • Specificity

    Detects Src only when phosphorylated at serine 75
  • Tested applications

    Suitable for: WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Dog, Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic phosphopeptide derived from Human Src around the phosphorylation site of serine 75 (VTSPPQ)

  • Positive control

    • IHC-P: Human brain tissue WB: extracts from COLO cells treated with EGF

Properties

Applications

Our Abpromise guarantee covers the use of ab79308 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 1/500 - 1/1000. Predicted molecular weight: 60 kDa.
ELISA 1/5000.
IHC-P 1/50 - 1/100.

Target

  • Function

    Non-receptor protein tyrosine kinase that plays pivotal roles in numerous cellular processes such as proliferation, migration, and transformation. In concert with PTK2B, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. Once it is recruited to the activated integrins, by PTK2B, it phosphorylates CBL which in turn induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. Phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
    Contains 1 SH3 domain.
  • Post-translational
    modifications

    Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation.
    S-nitrosylation is important for activation of its kinase activity.
  • Cellular localization

    Cell membrane. Mitochondrion inner membrane.
  • Information by UniProt
  • Database links

  • Form

    This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
  • Alternative names

    • ASV antibody
    • Avian sarcoma virus antibody
    • AW259666 antibody
    • c SRC antibody
    • CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA antibody
    • cSrc antibody
    • EC 2.7.10.2 antibody
    • Neuronal CSRC tyrosine specific protein kinase antibody
    • Neuronal proto-oncogene tyrosine-protein kinase Src antibody
    • Neuronal SRC antibody
    • Oncogene SRC antibody
    • OTTHUMP00000174476 antibody
    • OTTHUMP00000174477 antibody
    • p60 Src antibody
    • p60-Src antibody
    • p60c-src antibody
    • p60Src antibody
    • pp60c src antibody
    • pp60c-src antibody
    • pp60csrc antibody
    • Proto oncogene tyrosine protein kinase Src antibody
    • Proto-oncogene c-Src antibody
    • Proto-oncogene tyrosine-protein kinase Src antibody
    • Protooncogene SRC antibody
    • Protooncogene SRC Rous sarcoma antibody
    • Src antibody
    • SRC Oncogene antibody
    • SRC proto oncogene non receptor tyrosine kinase antibody
    • SRC_HUMAN antibody
    • SRC1 antibody
    • Tyrosine kinase pp60c src antibody
    • Tyrosine protein kinase SRC 1 antibody
    • Tyrosine protein kinase SRC1 antibody
    • v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog antibody
    • V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) antibody
    • v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian antibody
    see all

Images

  • All lanes : Anti-Src (phospho S75) antibody (ab79308) at 1/1000 dilution

    Lane 1 : extracts from COLO cells (treated
    with EGF, 200ng/ml, 30mins)
    Lane 2 : extracts from COLO cells (treated
    with EGF, 200ng/ml, 30mins) with immunizing peptide at 10 µg

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

  • Left panel: immunohistochemistry analysis of paraffin-embedded Human brain tissue with ab79308 at 1/100 dilution. Right panel: same, after pre-incubation with the immunizing peptide
  • ab79308 staining Src (phospho S75) in Dog osteosarcoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with acetone and blocked with 2% serum for 1 hour; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/50 in serum) for 14 hours at 4°C. A Biotinylated Goat anti-rabbit IgG (1/400) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:

  • Kashiwagi K  et al. A Ser75-to-Asp phospho-mimicking mutation in Src accelerates ageing-related loss of retinal ganglion cells in mice. Sci Rep 7:16779 (2017). Read more (PubMed: 29196663) »
  • Winter M  et al. Deciphering the Acute Cellular Phosphoproteome Response to Irradiation with X-rays, Protons and Carbon Ions. Mol Cell Proteomics 16:855-872 (2017). Read more (PubMed: 28302921) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (osteosarcoma)
Specification
osteosarcoma
Fixative
Acetone
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%

Abcam user community

Verified customer

Submitted Jun 18 2012

Answer


I am very pleased to hear you would like to accept our offer these antibodiesindog. This code will give you: 1 freeprimary antibodybefore the expiration date for each antibody tested. To redeem this offer, please submit an Abreview for dog for each of the antibodies tested and include the specific code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for your reply and the additional information which is very helpful. The fact that the total protein gives you the same band pattern might indicate that these bands could be real for your rat samples. Which antibody did you use to detect total Src? Rat is listed as "predicted to work with" on the datasheet which means that it is only predicted based on sequence homology for the immunogen. We have no data for rat samples so far and do not know if and how well it would work with this species. Does the treatment to inactivate Src give you the expected results (increase/decrease in signal)? The total protein should nevertheless remain unchanged as the total protein should not be affected by changes in phosphorylation. Do you see this as well? I'd be happy to issue you a testing discount retroactively since you are using the antibody on an untested species. The discount code will give you a free antibody on a future purchase after the rat data have been submitted as an Abreview. Here are more details on how this works: For UNTESTED species and/or applications, we have established a testing discount program. The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount. Please let me know if I shall send you a discount code. I look forward to hear back from you.

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Answer

Thank you for contacting us and for including the WB image. The band shown on our datasheet runs at about 55 to 60 kDa and was blocked with the immunizing peptide. Only human samples have so far been tested with this antibody. This antibody detects phospho-Src. What does your total protein sample look like? And have you tried any other sample (e.g. human sample or cell line)? What antibody dilution are you using? And what are the blocking conditions? I am not quite certain what the 85 kDa band could be. The protein can be potentially phosphorylated at multiple sites, and depending on the location of the phospho-site (e.g. in proximity to a Proline residue) the MW can sometimes shift to a higher MW than expected. But since we have not tried rat samples, I am not sure if this band pattern is sample specific or species specific or if the antibody gives extra (non-specific?) bands in rat samples. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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