Src [p Tyr416] Antibody (1246F) [Unconjugated]

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Src at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Src at approximately 62 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Src phosphorylated at Y416 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line treated with Pervanadate using Rabbit Anti-Human Phospho-Src (Y416) Monoclonal Antibody (Catalog # MAB2685) at 2 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Aberrant phosphorylation of lamin A by Src may cause nuclear dysmorphia.(A) MCF7 and MDA-MB-231 cells were treated with (+) or without (−) 50 nM dasatinib for 1 h and then lysed. Equal amounts of the whole cell lysates (WCLs) were incubated with anti-lamin A/C or pre-immune serum (IgG) as the control. The immunonocomplexes were analyzed by immunoblotting (IB) with anti-PY or anti-lamin A/C antibodies. An equal amount of WCLs was analyzed by immunoblotting with anti-Src, anti-Src pY416, or anti-actin. The tyrosine phosphorylation of lamin A was quantified and expressed as -fold relative to the level of MCF7 without dasatinib. (B) MCF7 and MDA-MB-231 cells were treated with (+) or without (−) 50 nM dasatinib for 1 h. The cells were fixed and stained for lamin A, lamin B1, and DNA. Representative images are shown. Scale bars, 10 μm. The nuclear circularity (4π × area/perimeter2) was determined. The P-values were calculated from at least 150 cells pooled from three independent experiments. The percentage of the cells with nuclear lobulation was measured (n ≥ 400). The values (mean ± SD) are from three experiments. ***P < 0.001. (C) MDA-MB-231 cells were infected with lentivirues capable of expressing FLAG-lamin A or the mutants (Y45F and Y45D) and selected in the medium with neomycin (neo). An equal amount of WCLs was analyzed by immunoblotting (IB) with antibodies as indicated. (D) The cells as described in panel C were fixed and stained for FLAG-lamin A, lamin B1, and DNA. Representative images are shown. Scale bars, 10 μm. The percentage of the cells with nuclear lobulation was measured (n ≥ 900). Values (means ± SD) are from three experiments. *P < 0.05, **P < 0.01, ***P < 0.001.Source data are available for this figure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34385357), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF
• Clone: 1246F
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

Src [p Tyr416] Antibody (1246F) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Phosphopeptide containing the human Src Y416 site
• Modification: p Tyr416
• Specificity: Detects human Src when phosphorylated at Y416 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: SRC
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1-25 ug/mL
  • Simple Western 1 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Src [p Tyr416] Antibody (1246F) [Unconjugated]

• ASV
• c-Src
• EC 2.7.10
• EC 2.7.10.2
• pp60c-src
• Rous sarcoma
• RSVgp4
• Src
• tyrosine kinase pp60c-src
• tyrosine-protein kinase SRC-1
• v-src avian sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog
• v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian)

Background

The Src family of proteins are intracellular tyrosine kinases involved in cell proliferation, differentiation, motility, and survival. Src family activity is regulated by tyrosine phosphorylation at two sites with opposing effects. Autophosphorylation in the activation loop of the kinase domain (Y419 of human Src) upregulates the enzyme, while phosphorylation in the C-terminal tail (Y530 of human Src) downregulates activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Src Antibody (MAB2685)(6)

Publications using MAB2685

• Yu-Hsuan Chen, Jinn-Yuan Hsu, Ching-Tung Chu, Yao-Wen Chang, Jia-Rong Fan, Muh-Hwa Yang et al. Loss of cell–cell adhesion triggers cell migration through Rac1-dependent ROS generation Life Science Alliance 2023-02-01 [PMID: 36446524] (Immunohistochemistry, Human)
• Spyridoula-Angeliki Nikou, Chunsheng Zhou, James S. Griffiths, Natalia K. Kotowicz, Bianca M. Coleman, Mary J. Green et al. The Candida albicans toxin candidalysin mediates distinct epithelial inflammatory responses through p38 and EGFR-ERK pathways Science Signaling 4/5/2022 [PMID: 35380879]
• J Chean, CJ Chen, G Gugiu, P Wong, S Cha, H Li, T Nguyen, S Bhattichar, JE Shively Human CEACAM1-LF regulates lipid storage in HepG2 cells via fatty acid transporter CD36 The Journal of Biological Chemistry, 2021-10-16;0(0):101311. 2021-10-16 [PMID: 34666041] (Western Blot, Human)
• Ching-Tung Chu, Yi-Hsuan Chen, Wen-Tai Chiu, Hong-Chen Chen Tyrosine phosphorylation of lamin A by Src promotes disassembly of nuclear lamina in interphase Life Science Alliance 10/1/2021 [PMID: 34385357]
• Seok-Young Kim, Sang-Min Kim, Sumin Lim, Ji Yeon Lee, Su-Jin Choi, San-Duk Yang et al. Modeling Clinical Responses to Targeted Therapies by Patient-Derived Organoids of Advanced Lung Adenocarcinoma Clinical Cancer Research 8/1/2021 [PMID: 34083237]
• L Basova, JA Najera, N Bortell, D Wang, R Moya, A Lindsey, S Semenova, RJ Ellis, MCG Marcondes Dopamine and its receptors play a role in the modulation of CCR5 expression in innate immune cells following exposure to Methamphetamine: Implications to HIV infection PLoS ONE, 2018-06-26;13(6):e0199861. 2018-06-26 [PMID: 29944719] (Western Blot, Human)

Bioinformatics

• Gene Symbol: SRC