Recombinant
RabMAb

Recombinant Anti-SRC Family (phospho Y418) antibody [EP503Y] - BSA and Azide free (ab239824)

Overview

  • Product name

    Anti-SRC Family (phospho Y418) antibody [EP503Y] - BSA and Azide free
    See all SRC Family primary antibodies
  • Description

    Rabbit monoclonal [EP503Y] to SRC Family (phospho Y418) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ELISA, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SRC Family aa 350-450 (phospho Y418). The exact sequence is proprietary.
    Database link: P07948

  • General notes

    ab239824 is the carrier-free version of ab40660 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab239824 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and SRC (P12931). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.

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

    This product is a recombinant rabbit monoclonal antibody.

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

    Constituent: PBS
  • Concentration information loading...
  • Clonality

    Monoclonal
  • Clone number

    EP503Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab239824 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. Detects a band of approximately 56 kDa (predicted molecular weight: 61 kDa).Can be blocked with Lyn (phospho Y396) peptide (ab202645).

Induction of cells with Pervanadate: Prepare 500mM stock Sodium Orthovanadate in PBS. To make 100mM (100x) Pervanadate Na3VO4 right before experiment, add 400ul 500mM Sodium Orthovanadate to 1560ul H2O and then 40ul H2O2, vortex and incubate at room RT for 5-10min. Cells are serum-starved overnight, and then treated with freshly made Pervanadate (100x, final concentration 1mM) for 20min at 37°C.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance

    SRC Family Negative Regulatory Phosphorylation Site belongs to the protein kinase superfamily, Tyr protein kinase family, SRC subfamily. It contains one protein kinase domain, one SH2 domain and one SH3 domain. It may serve as part of a signaling pathway coupling the Fc receptor to the activation of the respiratory burst and may also contribute to neutrophil migration and may regulate the degranulation process of neutrophils. SRC Family Negative Regulatory Phosphorylation Site is expressed predominantly in cells of the myeloid and B lymphoid lineages. There are two named isoforms.
  • Cellular localization

    Isoform 1: Membrane; Lipid anchor. Isoform 2: Membrane; Lipid anchor. Cytoplasm.
  • Database links

  • Alternative names

    • ASV antibody
    • Protooncogene SRC, Rous sarcoma antibody
    • SRC antibody
    • SRC1 antibody

Images

  • Flow Cytometry analysis of HeLa cells (treated with 1mM pervanadate for 30 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/80 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody.

    Black - Isotype control, rabbit monoclonal IgG with pervanade treated HeLa cells.

    Blue - Unlabelled control, pervanadate treated HeLa cells without incubation with primary and secondary antibodies.

    Red - Pervandate treated HeLa cells with ab40660.

    Green - Untreated HeLa cells labelled with ab40660.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40660).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells (untreated and treated with 1mM pervanadate for 15 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40660).

  • Direct ELISA antibody dose-response curve using unpurified ab40660 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40660).

  • ICC/IF image of unpurified ab40660 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab40660, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40660).

References

ab239824 has not yet been referenced specifically in any publications.

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