Recombinant Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP503Y] to SRC Family (phospho Y418)
- Suitable for: WB, Flow Cyt (Intra), ICC/IF, ELISA
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SRC Family (phospho Y418) antibody [EP503Y]
See all SRC Family primary antibodies -
Description
Rabbit monoclonal [EP503Y] to SRC Family (phospho Y418) -
Host species
Rabbit -
Specificity
This antibody recognises Lyn phosphorylation on Tyrosine 396.BLAST analysis show 100% homology with Hck (pY410) and Lck (pY393), and 92% homology with Src (pY418), Fyn (pY419) and Yes (pY425) Therefore, the antibody may cross-react with these proteins.
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Tested applications
Suitable for: WB, Flow Cyt (Intra), ICC/IF, ELISAmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- WB: HeLa and C6 cell lysates treated with pervanadate. ICC/IF: HeLa cells treated with pervanadate. Flow Cyt (intra): HeLa cells treated with pervanadate.
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General notes
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.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA, 49% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP503Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab201858)
- Alexa Fluor® 647 Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab201860)
- Alexa Fluor® 594 Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab206347)
- PE Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab208828)
- Anti-SRC Family (phospho Y418) antibody [EP503Y] - BSA and Azide free (ab239824)
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40660 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 |
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WB | (2) |
1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 61 kDa).
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 (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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ELISA |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 61 kDa). 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 (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
ELISA
Use at an assay dependent concentration. |
Target
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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
- Entrez Gene: 6714 Human
- Entrez Gene: 20779 Mouse
- Entrez Gene: 83805 Rat
- Omim: 190090 Human
- SwissProt: P12931 Human
- SwissProt: P05480 Mouse
- SwissProt: Q9WUD9 Rat
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Alternative names
- ASV antibody
- Protooncogene SRC, Rous sarcoma antibody
- SRC antibody
- SRC1 antibody
Images
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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).
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Intracellular 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 HeLacells.
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.
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All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/2000 dilution (purified)
Lane 1 : Untreated HeLa whole cell lysate
Lane 2 : Pervanadate treated HeLa whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 56-59 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and dilution buffer: 5% BSA/TBST.
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All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)
Lane 1 : Pervanadate treated Jurkat cell lysate with Lyn non-phospho peptide
Lane 2 : Pervanadate treated Jurkat cell lysate with Lyn (pY396) phospho peptide
Lane 3 : Pervanadate treated Jurkat cell lysate with Lck non-phospho peptide
Lane 4 : Pervanadate treated Jurkat cell lysate with Lck (pY393) phospho peptide
Lane 5 : Pervanadate treated Jurkat cell lysate with Src non-phospho peptide
Lane 6 : Pervanadate treated Jurkat cell lysate with Src (pY418) phospho peptide
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 56-59 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)
Lane 1 : Untreated C6 whole cell lysate
Lane 2 : Pervanadate treated C6 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 56-59 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking and dilution buffer: 5% BSA/TBST.
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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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (39)
ab40660 has been referenced in 39 publications.
- Zhang H et al. QKI-6 Suppresses Cell Proliferation, Migration, and EMT in Non-Small Cell Lung Cancer. Front Oncol 12:897553 (2022). PubMed: 35600368
- Zhang J et al. Na/K-ATPase suppresses LPS-induced pro-inflammatory signaling through Lyn. iScience 25:104963 (2022). PubMed: 36072548
- Zeng W et al. Lidocaine suppresses the malignant behavior of gastric cancer cells via the c-Met/c-Src pathway. Exp Ther Med 21:424 (2021). PubMed: 33747163
- Rip J et al. A Versatile Protocol to Quantify BCR-mediated Phosphorylation in Human and Murine B Cell Subpopulations. Bio Protoc 11:e3902 (2021). PubMed: 33732789
- Kawano M et al. Analysis of the signal cross talk via CCL26 in the tumor microenvironment in osteosarcoma. Sci Rep 11:18099 (2021). PubMed: 34518591