Human Phospho-Src (Y416) Antibody MAB2685: R&D Systems

Human Phospho-Src (Y416) Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Src when phosphorylated at Y416 in direct ELISAs and Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1246F
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Phosphopeptide containing the human Src Y416 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Simple Western
    1 µg/mL
    See below
  • Immunocytochemistry
    1-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Src by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Src at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Src by Simple WesternTM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Src at approximately 62 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Immunocytochemistry
Phospho-Src (Y416) in HepG2 Human Cell Line. Src phosphorylated at Y416 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line treated with Pervanadate using Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685) at 2 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Src

The Src family of proteins are intracellular tyrosine kinases involved in cell proliferation, differentiation, motility, and survival. Src family activity is regulated by tyrosine phosphorylation at two sites with opposing effects. Autophosphorylation in the activation loop of the kinase domain (Y419 of human Src) upregulates the enzyme, while phosphorylation in the C-terminal tail (Y530 of human Src) downregulates activity.

  • Long Name:
    v-src Sarcoma [Schmidt-Ruppin A-2] Viral Oncogene Homolog
  • Entrez Gene IDs:
    6714 (Human); 20779 (Mouse); 83805 (Rat); 1491925 (Viral)
  • Alternate Names:
    ASV; c-Src; EC 2.7.10; EC 2.7.10.2; pp60c-src; Rous sarcoma; RSVgp4; Src; tyrosine kinase pp60c-src; tyrosine-protein kinase SRC-1; v-src avian sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog; v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian)
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