RabMAb

Anti-SRC Family (phospho Y418)抗体[EP503Y] (ab40660)

Abreviews (1)Q&A (6)Specific References (12)
Customer reviews and FAQs

概述

  • 产品名称Anti-SRC Family (phospho Y418)抗体[EP503Y]
    参阅全部 SRC Family 一抗
  • 描述
    兔单克隆抗体[EP503Y] to SRC Family (phospho Y418)
  • 特异性ab40660 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, athe antibody may cross-react with these proteins.
  • 经测试应用适用于: WB, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human SRC Family aa 350-450 (phospho Y418). The exact sequence is proprietary.
    Database link: P07948
    (Peptide available as ab202645)

  • 阳性对照
    • WB: HeLa and C6 cell lysates treated with pervanadate. ICC/IF: HeLa cells treated with pervanadate. Flow Cyt: HeLa cells treated with pervanadate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.
    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    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.

    Alternative versions available:
    Anti-Lyn (phospho Y396) antibody (Alexa Fluor® 594) [EP503Y] (ab206347)

     

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • 克隆单克隆
  • 克隆编号EP503Y
  • 同种型IgG
  • 研究领域

应用

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.

应用 Ab评论 说明
WB 1/1000 - 1/10000. 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 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.

靶标

  • 相关性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.
  • 细胞定位Isoform 1: Membrane; Lipid anchor. Isoform 2: Membrane; Lipid anchor. Cytoplasm.
  • 数据库链接
    see all
  • 别名
    • ASV antibody
    • Protooncogene SRC, Rous sarcoma antibody
    • SRC antibody
    • SRC1 antibody

Anti-SRC Family (phospho Y418) antibody [EP503Y] 图像

  • Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    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
    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 seconds

    Blocking and dilution buffer: 5% BSA/TBST.

  • Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    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
    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 second

    Blocking and dilution buffer: 5% BSA/TBST.

  • Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    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
    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 minutes

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (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).

  • Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (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.

  • Flow Cytometry - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    Flow Cytometry - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

    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.

  • ELISA - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)
    ELISA - Anti-SRC Family (phospho Y418) antibody [EP503Y] (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.

Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)参考文献

This product has been referenced in:
  • Chiricozzi E  et al. Direct interaction, instrumental for signaling processes, between LacCer and Lyn in the lipid rafts of neutrophil-like cells. J Lipid Res 56:129-41 (2015). Read more (PubMed: 25418321) »
  • Nagao T  et al. Proliferation and survival signaling from both Jak2-V617F and Lyn involving GSK3 and mTOR/p70S6K/4EBP1 in PVTL-1 cell line newly established from acute myeloid leukemia transformed from polycythemia vera. PLoS One 9:e84746 (2014). Read more (PubMed: 24404189) »

See all 12 Publications for this product

发表研究结果有使用 ab40660？请让我们知道，以便我们可以引用本数据表中的参考文章。

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Western blot abreview for Anti-Lyn (phospho Y396) antibody [EP503Y]

 Good
Abreviews
Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (10% PAGE Gel)
Sample Human Cell lysate - whole cell (Hela cells)
Specification Hela cells
Treatment 100ng/ml IL6 for 15 minutes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Abcam user community

Verified customer

提交于 Feb 12 2015

One of our customers has ordered ab40660 and receives two bands close in size on the blot.
Does the antibody recognize Lyn a and Lyn b? that could explain it..
Is the epitope recognized by the antibody common to Lyna and Lyn b?

The difference between lyn a and lyn b is that lyn b does not contain amino acids 23-43 of lyn a. Since this antibody recognises a region around tyrosine 396, ab40660 can detect both lyn a and lyn b therefore it is very probable that this doublet band ...

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Inquiry: Hi Abcam I don't have a problem with this product but I need some info on it: please can you find out if this antibody detects pLyn(396) ONLY, or does it detect phosphorylation of any other src family member site? thankyou for your help

Thank you for your enquiry regarding ab40660.
Based on the sequence homology, it is unlikely that this antibody would cross react with the other tyrosines of Lyn. However, it might cross-react with LCK, HCK and possibly Src when phosphorylated at ...

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Please send the IgG concentrations of the following lots.
ab33906 lot YH090207C -
ab33862 lot YH082410D -
ab32429 lot YG080503 -
ab40660 lot YI070802CS -
ab32445 lot YI072804DS -

ab33906 lot YH090207C - 0.3260 mg/ml

ab33862 lot YH082410D - 0.6380 mg/ml

ab32429 lot YG080503 - 0.0790 mg/ml

ab40660 lot YI070802CS - 0.1030 mg/ml

ab32445 lot YI072804DS - 0.0250 mg/ml

Hi Abcam,

Could you please advise for the following enquiry;

I have a question regarding the cross reactivity of this Ab. There is contradicting information on the Abcam and Epitomics websites. Whereas the data sheet on the Epitom...

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Thank you for your enquiry.

I can confirm that we have a dedicated team at Abcam who review the literature in order to find publications that have used our products. In this case, the following publication was found, showing use of this anti...

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I recently purchased the products ab40660 (Lyn-pY396) and ab33914 (Lyn -pY507), but could you tell me the antibody stock concentrations please?

Thank you for your enquiry.

These 2 antibodies have lot specific concentrations. If you could please forward me the lot numbers for ab40660 and ab33914 I'd be happy to enquire about the concentrations.

Thanks for the additional info...

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I'd just like you to confirm/show the datawhere the Lyn pY396 antibody (Catalog No. ab40660) cross reacts with mouse Lyn? The reason I ask is that other companies selling the exact same clone (EP503Y) state that the antibody cross reacts with human Lyn...

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Thank you for contacting Abcam.

We added the reactivity with mouse based on the following reference:

Kim SW et al. Rational combined targeting of phosphodiesterase 4B and SYK in DLBCL. Blood 113:6153-60 (2009).



...

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