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  1. Link

    src-antibody-egtr103-ab133283.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Src Family
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Src antibody [EGTR103] (ab133283)

  • Datasheet
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Western blot - Anti-Src antibody [EGTR103] (ab133283)
  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
  • Anti-Src antibody [EGTR103] (ab133283)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EGTR103] to Src
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Src antibody [EGTR103]
    See all Src primary antibodies
  • Description

    Rabbit monoclonal [EGTR103] to Src
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Src aa 350-450.
    Database link: P12931

  • Positive control

    • IHC-P: Rat and Mouse colon tissues. Human liver tissue. WB: HAP1, A431, U87-MG, and SH-5YSY cell lysates. Mouse and rat brain lysate. Mouse hippocampus lysate
  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EGTR103
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Src Family
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Proto-oncogenes
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • Src family
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Src antibody [EGTR103] - BSA and Azide free (ab222221)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human Src protein (ab79635)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133283 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-P
1/50.

See IHC antigen retrieval protocols.

WB
1/1000. Predicted molecular weight: 60 kDa.

For unpurified use at 1/1000 - 1/10000.

Notes
IHC-P
1/50.

See IHC antigen retrieval protocols.

WB
1/1000. Predicted molecular weight: 60 kDa.

For unpurified use at 1/1000 - 1/10000.

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.
  • Target information above from: UniProt accession P12931 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6714 Human
    • Entrez Gene: 20779 Mouse
    • Entrez Gene: 83805 Rat
    • Omim: 190090 Human
    • SwissProt: P00523 Human
    • SwissProt: P12931 Human
    • SwissProt: P05480 Mouse
    • SwissProt: Q9WUD9 Rat
    • Unigene: 195659 Human
    • Unigene: 112600 Rat
    see all
  • 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

  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Western blot - Anti-Src antibody [EGTR103] (ab133283)
    All lanes : Anti-Src antibody [EGTR103] (ab133283) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : SRC knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 60 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-Src antibody [EGTR103] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133283 was shown to bind specifically to Src. A band was observed at 65 kDa in wild-type HAP1 cell lysates with no signal observed at this size in SRC knockout cell line HAP1. To generate this image, wild-type and SRC knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Lanes 1-3 : Anti-Src antibody [EGTR103] (ab133283) at 1/1000 dilution (Purified)
    Lane 4 : Purified at 1/1000 dilution

    Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
    Lane 2 : Mouse brain lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Mouse hippocampus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Anti-Src antibody [EGTR103] (ab133283) at 1/2000 dilution + U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat colon tissue sections labeling Src with purified ab133283 at 1/50 dilution (6.98 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse colon tissue sections labeling Src with purified ab133283 at 1/50 dilution (6.98 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Src antibody [EGTR103] (ab133283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling Src with purified ab133283 at 1/50 dilution (6.98 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Western blot - Anti-Src antibody [EGTR103] (ab133283)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Src knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A431 knockout HAP1 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab133283 observed at 60 kDa. Red - loading control, ab8226, observed at 42 kDa.
    Unpurified ab133283 was shown to specifically react with Src when Src knockout samples were used. Wild-type and Src knockout samples were subjected to SDS-PAGE. ab133283 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW)preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-Src antibody [EGTR103] (ab133283)
    Western blot - Anti-Src antibody [EGTR103] (ab133283)
    All lanes : Anti-Src antibody [EGTR103] (ab133283) at 1/1000 dilution (Unpurified)

    Lane 1 : U87-MG cell lysate
    Lane 2 : SH-5YSY cell lysate

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 60 kDa

  • Anti-Src antibody [EGTR103] (ab133283)
    Anti-Src antibody [EGTR103] (ab133283)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (3)

Publishing research using ab133283? Please let us know so that we can cite the reference in this datasheet.

ab133283 has been referenced in 3 publications.

  • Apaya MK  et al. Deregulating the CYP2C19/Epoxy-Eicosatrienoic Acid-Associated FABP4/FABP5 Signaling Network as a Therapeutic Approach for Metastatic Triple-Negative Breast Cancer. Cancers (Basel) 12:N/A (2020). PubMed: 31941087
  • Yang CC  et al. Plant galactolipid dLGG suppresses lung metastasis of melanoma through deregulating TNF-a-mediated pulmonary vascular permeability and circulating oxylipin dynamics in mice. Int J Cancer 143:3248-3261 (2018). PubMed: 29978476
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371

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