SRC1 antibody

Principal name

SRC1 antibody

Alternative names for SRC1 antibody

c-Src, pp60c-src, Proto-oncogene tyrosine-protein kinase Src, Proto-oncogene c-Src

SwissProt ID

P00523 (Chick), P05480 (Mouse), P12931 (Human), Q9WUD9 (Rat)

Gene ID

6714 (SRC), 20779 (Src)

Available reactivities

Hu (Human), Ms (Mouse), Rt (Rat), Can (Canine), Mky (Monkey), Av (Birds), Hst (Hamster), Chk (Chicken), Frog, Bov (Bovine), GP (Guinea Pig), Por (Porcine), Rb (Rabbit), Sh (Sheep), Trout, Xen (Xenopus), Ze (Zebrafish), Vertebrates, Ve

Available hosts

Rabbit, Mouse

Available applications

Immunoprecipitation (IP), Western blot / Immunoblot (WB), Enzyme Immunoassay (E), Immunocytochemistry/Immunofluorescence (ICC/IF), Flow Cytometry (F), Frozen Sections (C), Paraffin Sections (P), Radioimmunoassay (R), Dot blot (Dot), ELISA (detection) (E(detection))

Background of SRC1 antibody

c-Src tyrosine kinase plays a critical role in signal transduction downstream of growth factor receptors, integrins and G protein-coupled receptors. We used stable isotope labeling with amino acids in cell culture (SILAC) approach to identify additional substrates of c-Src tyrosine kinase in human embryonic kidney 293T cells. We have identified 10 known substrates and interactors of c-Src and Src family kinases along with 26 novel substrates. We have experimentally validated 4 of the novel proteins (NICE-4, RNA binding motif 10, FUSE-binding protein 1 and TRK-fused gene) as direct substrates of c-Src using in vitro kinase assays and cotransfection experiments. Significantly, using a c-Src specific inhibitor, we were also able to implicate 3 novel substrates (RNA binding motif 10, EWS1 and Bcl-2 associated transcription factor) in PDGF signaling. Finally, to identify the exact tyrosine residues that are phosphorylated by c-Src on the novel c-Src substrates, we designed custom peptide microarrays containing all possible tyrosine-containing peptides (312 unique peptides) and their mutant counterparts containing a Tyr --> Phe substitution from 14 of the identified substrates. Using this platform, we identified 34 peptides that are phosphorylated by c-Src. We have demonstrated that SILAC-based quantitative proteomics approach is suitable for identification of substrates of nonreceptor tyrosine kinases and can be coupled with peptide microarrays for high-throughput identification of substrate phosphopeptides.

General readings

Western Blot
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a transfected cell lysate sample (cell lysis buffer: 50 mM Tris-HCl, pH 7.4; 1% NP-40; 0.25% sodium deoxycholate; 150 mM NaCl; 1 mM EDTA; 1 mM PMSF; 1 μg/mL each aprotinin, leupeptin, pepstatin; 1 mM Na3VO4, 1 mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water.
2. Block the blotted nitrocellulose in freshly prepared 3% nonfat dry milk in TBS with 0.05% Tween®-20 (TBST-MLK) for 1 hour at room temperature with constant agitation.
3. Incubate the nitrocellulose with a 1:1,000 dilution of anti-phospho-Src family (Tyr416), clone 2N8, in freshly prepared TBST-MLK overnight with agitation at 4°C.
4. Wash the nitrocellulose twice with water.
5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-rabbit HRP conjugated IgG 1:5000 dilution was used) in TBST-MLK for 1 hour with agitation at room temperature.
6. Wash the nitrocellulose twice with water.
7. Wash the nitrocellulose in TBS-0.05% Tween®-20 for 3-5 minutes.
8. Rinse the nitrocellulose in 4-5 changes of water.
9. Use detection method of choice (enhanced chemiluminescence was used).

Immunoprecipitation Protocol for detection of endogenous phospho-Src (Tyr416)

1. Dilute the sample cell lysate to roughly 1 μg/μL total cell protein with PBS.
2. Add 500 μg -1 mg of the cell lysate to a 1.5 mL microfuge tube.
3. Add 30 μL of anti-Src agarose, clone GD11 beads.
4. Gently rock the reaction overnight at 4°C.
5. Collect the agarose beads by pulsing (5 seconds in the microcentrifuge at 14,000 x g), and drain off the supernatant. Wash the beads 3 times with either ice-cold cell lysis buffer or PBS.
6. Resuspend the agarose beads in 60 μL of 2X Laemmli sample buffer.
7. Store the beads frozen for future analysis or boil the beads for 5 minutes.
8. Collect the beads after boiling using a microcentrifuge pulse.
9. Perform SDS-PAGE and immunoblot analysis on a sample of the supernatant fraction using anti-phospho-Src family (Tyr416), clone 2N8 as the primary antibody.

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

SRC1 antibody

Immunohistochemical analysis of paraffin-embedded bladder cancer tissues using SRC Mouse mAb with DAB staining. Mouse IgG1 1F11 Ascites Hu E, F, P, WB
0.1 ml / €430.00
  Acris Antibodies GmbH

SRC1 antibody

Western blot analysis of extracts from HeLa cells, using Src (Ab-529) Antibody.The lane on the right is treated with the synthesized peptide. Rabbit IgG Purified Hu, Ms, Rt E, P, WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 antibody

Predicted band size: 60 kDa . Positive control: PC3 cell lysate. Recommended dilution: 1/200-1000. (Gel: 10%SDS-PAGE Lysate: 40 ug per lane Primary antibody: 1/100 dilution Secondary antibody: Goat anti Rabbit IgG - H&L (HRP) at 1/10000 dilution Exposure time: 1 minute) Rabbit IgG Purified Hu, Ms, Rt E, P, WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 antibody

Predicted band size: 60 kDa . Positive control: PC3 cell lysate. Recommended dilution: 1/200-1000. (Gel: 10%SDS-PAGE Lysate: 40 ug per lane Primary antibody: 1/200 dilution Secondary antibody: Goat anti Rabbit IgG - H&L (HRP) at 1/10000 dilution Exposure time: 2 minutes) Rabbit IgG Purified Hu, Ms, Rt E, P, WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 antibody

Western blot analysis of Src expression in 293 whole cell lysates,The lane on the left is treated with the antigen-specific peptide. Rabbit IgG Purified Hu, Ms, Rt WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 antibody

Western blot analysis of Src expression in COLO205 whole cell lysates,The lane on the left is treated with the antigen-specific peptide. Rabbit IgG Purified Hu, Ms, Rt WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 antibody

Western blot analysis of Src expression in HeLa whole cell lysates,The lane on the left is treated with the antigen-specific peptide. Rabbit IgG Purified Hu, Ms, Rt WB
0.1 ml / €325.00
  OriGene Technologies, Inc.

SRC1 pTyr416 antibody

SRC1 Mouse IgG3 9A6 Biotin Hu, Ms E, F, ICC/IF, WB
0.1 mg / €490.00
  Acris Antibodies GmbH

SRC1 pTyr416 antibody

Phosphospecificity: Whole cell extracts of control (co) and pervanadate stimulated HEK-293 tumor cells were applied to SDS-PAGE (ca 20.000 cells per lane) and transferred to PVDF membranes. Immunoblots were probed with mab src 9A6 (0.5 μg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec). lane 1: co; lane2: pervanadate Mouse IgG3 9A6 Purified Hu, Ms E, F, ICC/IF, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

SRC1 (N-term) (incl. pos. control) antibody

Detection of endogenous Src: Whole cell extracts of vanadate treated tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. 
The immunoblot was probed with AM00146PU-N Src antibody Clone 11F1 (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec).
Lane 1: A431
Lane 2: SW480
Lane 3: SW620
Lane 4: HT29
Lane 5: MCF-7
Lane 6: MDA-MB231
Lane 7: T47D Mouse IgG1 11F1 Purified Can, Hu, Ms, Rt E, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

SRC1 antibody

Western blot analysis using SRC antibody Cat.-No AM06226SU-N against truncated SRC-His recombinant protein (Lane 1) and PMA treated THP-1 cell lysate (Lane 2). Mouse IgG2b 4F1E8 Ascites Hu E, WB
0.1 ml / €460.00
  Acris Antibodies GmbH

SRC1 antibody

Western blot analysis using SRC mouse mAb against truncated SRC-His recombinant protein (1) and PMA treated THP-1 cell lysate (2). Mouse IgG2a 5D10C4 Ascites Hu WB
0.1 ml / €440.00
  Acris Antibodies GmbH

SRC1 antibody

Figure 1. Western blot analysis using anti-Src Mab (Cat. #AM11074PU-N) to detect Src in Jurkat cell lysate. Mouse IgG1 17AT28 Purified Hu, Ms IP, P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

SRC1 (full length) antibody

AM20333PU-N SRC antibody staining of Formalin-Fixed,  Paraffin-Embedded Human Adrenal at 20 µg/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen. Mouse 5A18 Purified Hu, Ms, Rt P, WB
50 µg / €470.00
  Acris Antibodies GmbH

SRC1 pTyr418 antibody

Immunohistochemistry (IHC) analyzes of Src pTyr418 antibody (Cat.-No.: AP01687PU-N) in paraffin-embedded human colon carcinoma tissue. Rabbit Aff - Purified Hu, Ms, Rt P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

SRC1 pTyr529 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Src (phospho-Tyr529) antibody AP02437PU. Rabbit Aff - Purified Hu P, WB
0.1 ml / €320.00
  Acris Antibodies GmbH

SRC1 pTyr529 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Src (phospho-Tyr529) antibody AP02437PU. Rabbit Aff - Purified Hu P, WB
50 µl / €240.00
  Acris Antibodies GmbH

SRC1 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Src antibody AP02687PU. Rabbit Aff - Purified Hu, Ms, Rt P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

SRC1 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Src antibody AP02687PU. Rabbit Aff - Purified Hu, Ms, Rt P, WB
50 µl / €230.00
  Acris Antibodies GmbH

SRC1 antibody

Western blot analysis using Src [pY529] antibody on CEF cells untransfected (1) or transfeceted with wild-type human Src (2-5). Antibody was incubated prior to use with no peptide (1,2), nonphosphorylated immunogen peptide (3), generic phosphotyrosine containing peptide (4), or phosphorylated immunogen peptide (5). Rabbit Purified Chk, Frog, Hu, Ms, Rt C, WB
0.1 ml / €710.00
  Acris Antibodies GmbH

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