Name: Anti-SRC Family (phospho Y418) antibody [EP503Y]
Brand: RabMAb
SKU: ab40660
Rating: 4 (2 reviews)

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Immunocytochemistry/ Immunofluorescence - 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

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free PE

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)
宿主

Rabbit
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, the antibody may cross-react with these proteins.
经测试应用

适用于: WB, Flow Cyt (Intra), ICC/IF, ELISAmore 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 (intra): HeLa cells treated with pervanadate.
常规说明
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
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.

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

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA, 49% PBS
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EP503Y
同种型

IgG
研究领域

The Abpromise guarantee

Abpromise™承诺保证使用ab40660于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.
ICC/IF		Use at an assay dependent concentration.
ELISA		Use at an assay dependent concentration.

说明
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.

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
- Entrez Gene: 6714 Human
- Entrez Gene: 20779 Mouse
- Entrez Gene: 83805 Rat
- Omim: 190090 Human
- SwissProt: P12931 Human
- SwissProt: P05480 Mouse
- SwissProt: Q9WUD9 Rat
别名
- ASV antibody
- Protooncogene SRC, Rous sarcoma antibody
- SRC antibody
- SRC1 antibody

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

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.
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
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 seconds

Blocking and dilution buffer: 5% BSA/TBST.
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
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 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
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
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 second

Blocking and dilution buffer: 5% BSA/TBST.
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.
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)

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.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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。

ab40660 被引用在 39 文献中.

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

View all Publications for this product

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1-8 of 8 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (U2OS)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

U2OS

Blocking step

Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 30 2018

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 12 2015

Question

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?

Abcam community

Verified customer

Asked on Feb 24 2014

Answer

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 seen at around 57 kDa corresponds to a band at 56 kDa for lyn b and 59 kDa for lyn a.

Elisa Thomas

Abcam Scientific Support

回复于 Feb 24 2014

Question

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

Abcam community

Verified customer

Asked on Mar 06 2013

Answer

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 equivalent sites since Lyn shares high sequence homology with other Src family members.
Please note that the development of all phospho-specific antibodies requires to screen on phospho specific peptide and counter-screen on its corresponding non-phospho specific control peptide.
If you need any further assistance in the future, please do not hesitate to contact me.

Abcam Scientific Support

回复于 Mar 06 2013

Question

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

Abcam community

Verified customer

Asked on Feb 05 2013

Answer

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

Abcam Scientific Support

回复于 Feb 05 2013

Question

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 Epitomics website says the Ab does not recognise murine Lyn, the Abcam product data sheet reports cross reactivity with murine Lyn.

Could you please clarify.

Thanks for your help!

Kind regards,

Abcam community

Verified customer

Asked on Oct 30 2012

Answer

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 antibody in mouse. This link is available on the datasheet:

Kim SW et al. Rational combined targeting of phosphodiesterase 4B and SYK in DLBCL. Blood 113:6153-60 (2009). WB; Mouse. PubMed: 19369227

Therefore, mouse has been added to the tested species list for ab40660 Lyn (phospho Y396) antibody [EP503Y].

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Abcam Scientific Support

回复于 Oct 30 2012

Question

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

Abcam community

Verified customer

Asked on Feb 17 2012

Answer

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 information!

Abcam Scientific Support

回复于 Feb 17 2012

Question

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, with no mention of mouseLynso I'd just like to be sure before ordering.

Abcam community

Verified customer

Asked on Feb 08 2012

Answer

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).

The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot onmouse samples . If you do experience any problems with it and we cannot resolve the issue, thenwe would be happy to either send a replacement antibody or to process a refund.

Please let me know if there is anything else I can help you with.

Abcam Scientific Support

回复于 Feb 08 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

应用

The Abpromise guarantee

靶标

相关性

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (39)

客户评价及客户问答

Western blot abreview for Anti-SRC Family (phospho Y418) antibody [EP503Y]

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