Antibodies to Focal Adhesion Kinase (FAK) - FocusOn 101

Introduction

Focal Adhesion Kinase (FAK) is a 125 kDa non-receptor protein tyrosine kinase involved in integrin-mediated control of cell behaviour.

Integrins are the major cell surface receptors for extracellular matrix molecules, which play critical roles in a variety of biological processes. Focal adhesion kinase (FAK) has been discovered as a substrate for Src and it is a key protein in the signal transduction pathways triggered by integrins. FAK plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. Aggregation of FAK with integrins and cytoskeletal proteins in focal contacts has been proposed to be responsible for FAK activation and autophosphorylation by integrins in cell adhesion.

Interaction with integrin and focal adhesion kinase (FAK) regulates the cancer cell adhesion and invasion into extracellular matrix (ECM). In addition, phosphorylation of FAK correlates with the increase of cell motility and invasion. Adhesion and spreading of cancer cells on a variety of ECM proteins, including collagen type IV (Coll IV), leads to an increase in tyrosine phosphorylation and activation of FAK. FAK regulation includes phosphorylation at multiple tyrosine and serine residues. Phosphorylation of tyrosine generally is associated with positive regulation and growth promotion, however, dephosphorylation at these sites occurs as cells enter mitosis (M-Phase of the cell cycle).

In contrast, serine phosphorylation either remains high or is increased as cells enter mitosis and may play a role in focal adhesion disassembly.

Antibody Panel to Focal Adhesion Kinase (FAK)

Acris Antibodies offers a range of polyclonal antibodies to FAK and phoshorylated FAK (serine or tyrosine phosphospecific antibodies). All products are suitable for Western blot application, some of these antibodies can be used for Immunohistochemistry and / or Immunofluorescence (for more details see pictures and table below.)

Peptide competition experiment using anti-FAKpSer732-antibody Cat. No. BP7066: <br><br>Extracts prepared from PC12 cells treated with nocozadole were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with FAK [pSer732] antibody for two hours at room temperature in a 1% Milk-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phospho-serine containing peptide (3), or, the phosphopeptide immunogen (4).<br><br>After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. <br><br>The data show that only the peptide corresponding to FAK [pSer732] blocks the antibody signal, thereby demonstrating the specificity of the antibody.Peptide competition experiment using anti-FAKpTyr861 antibody Cat. No. BP7073: <br><br>Extracts prepared from CEF cells expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with the FAK [pTyr861] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: (1) the phosphopeptide immunogen, (2) a generic phosphotyrosine containing peptide, (3) the non-phosphorylated peptide corresponding to the phosphopeptide, and (4) no peptide.<br><br>After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method.<br><br>The data show that only the peptide corresponding to FAK [pTyr861] blocks the antibody signal, thereby demonstrating the specificity of the antibody.

Fig.1: Peptide competition experiment using anti-FAKpSer732-antibody Cat. No. BP7066:

Extracts prepared from PC12 cells treated with nocozadolewere resolved by SDS-PAGE on a 10% polyacrylamide geland transferred to PVDF. Membranes were blocked with a5% BSA-TBST buffer for one hour at room temperature, thenincubated with FAK [pSer732] antibody for two hours at roomtemperature in a 1% Milk-TBST buffer, following priorincubation with: no peptide (1), the non-phosphopeptidecorresponding to the immunogen (2), a generic phosphoserinecontaining peptide (3), or, the phosphopeptide immunogen (4).After washing, membranes were incubated with goat F(ab’)2anti-rabbit IgG HRP conjugate and bands were detectedusing the Pierce SuperSignal method.

The data show that only the peptide corresponding to FAK[pSer732] blocks the antibody signal, thereby demonstratingthe specificity of the antibody.

Fig.2:Peptide competition experiment using anti-FAKpTyr861antibody Cat. No. BP7073:

Extracts prepared from CEF cells expressing FAK and plated onfibronectin were resolved by SDS-PAGE on a 10% polyacrylamidegel and transferred to PVDF. Membranes were blocked with a 5%BSA-TBST buffer overnight at 4°C, then were incubated with theFAK [pTyr861] antibody for two hours at room temperature in a 3%BSA-TBST buffer, following prior incubation with: (1) thePhosphopeptide immunogen, (2) a generic phosphotyrosinecontaining peptide, (3) the non-phosphorylated peptidecorresponding to the phosphopeptide, and (4) no peptide.

After washing, membranes were incubated with goat F(ab’)2 antirabbit IgG HRP conjugate and bands were detected using thePierce SuperSignal method.

The data show that only the peptide corresponding to FAK[pTyr861] blocks the antibody signal, thereby demonstrating thespecificity of the antibody.

Antibody Panel to FAK

Focal Adhesion Kinase (FAK) –antibody.

Literature

Sawai H, Okada Y, Funahashi H, Matsuo Y, Takahashi H, Takeyama H, Manabe T.Activation of focal adhesion kinase enhances the adhesion and invasion of pancreatic cancer cells viaextrcellular signal-regulated kinase-1/2 signaling pathway activation.Molecular Cancer 2005; 4:37.

Eliceiri, B.P., et al. (2002)Src-mediated coupling of focal adhesion kinase to integrin alpha(v)beta5 in vascular endothelial growthfactor signaling. J. Cell Biol. 157(1):149-160.

Hauck, C.R., et al. (2002)v-Src SH3-enhanced interaction with focal adhesion kinase at beta 1 integrin-containing invadopodiapromotes cell invasion. J. Biol. Chem. 277(15):12487-12490.

Guan JL.Role of focal adhesion kinase in integrin signalling.Int J Biochem Cell Biol. 1997 Aug-Sep; 29(8-9): 1085-96.

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Primary Antibodies

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

FAK1 / PTK2 pTyr397 (incl. pos. control) antibody

Detection of endogenous FAK: Whole cell lysates of serum starved tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to PVDF membranes. Immunoblots were probed with mab AM00059PU-N (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec).
Lane 1: HeLa 
Lane 2: HepG2 
Lane 3: HEK293 
Lane 4: SH-SY5Y 
Lane 5: MDCK 
Lane 6: PC12 
Lane 7: CMT 93 
Lane 8: Neuro 2A
Lane 9: 3T3 Mouse IgG1 2D11 Purified Can, Hu, Ms, Rt E, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

HeLa cells (lane 1), mouse L929 cells (lane 2), and rat PC12 cells (lane 3) were resolved by SDS-PAGE and transferred to PVDF. The membranes were incubated with this FAK monoclonal antibody (clone 34Q36) at a concentration of 1 μg/mL for two hours at room temperature. After washing, the membranes were incubated with a goat F(ab’)2 anti-mouse IgG alkaline phosphatase conjugated antibody at a 1:2000 dilution. Bands were detected with CDP-substrate using the WesternStarTM method (Tropix) and Kodak BioMax film. Mouse IgG2b 34Q36 Purified Hu, Ms, Rt WB
0.1 mg / €540.00
  Acris Antibodies GmbH

FAK1 / PTK2 (1-423) antibody

Tonsil: Formalin-Fixed Paraffin-Embedded (FFPE) Mouse IgG1 4.47 Purified Hu, Ms, Rt E, ICC/IF, IP, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

FAK1 / PTK2 Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, IP, WB
0.1 mg / €390.00
  Acris Antibodies GmbH

FAK1 / PTK2 pTyr861 antibody

Figure 1. Western blot analysis using FAK  antibody (Lane 1 and 2) and FAK (phospho-Tyr861) antibody AP02358PU (Lane 3, 4, 5 and 6). Rabbit Aff - Purified Hu, Ms WB
0.1 ml / €320.00
  Acris Antibodies GmbH

FAK1 / PTK2 pTyr861 antibody

Figure 1. Western blot analysis using FAK  antibody (Lane 1 and 2) and FAK (phospho-Tyr861) antibody AP02358PU (Lane 3, 4, 5 and 6). Rabbit Aff - Purified Hu, Ms WB
50 µl / €240.00
  Acris Antibodies GmbH

FAK1 / PTK2 pTyr925 antibody

Figure 2. Immunofluorescence staining of methanol-fixed HeLa cells using FAK (phospho-Tyr925) antibody (#AP02411PU, Red). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
0.1 ml / €320.00
  Acris Antibodies GmbH

FAK1 / PTK2 pTyr925 antibody

Figure 2. Immunofluorescence staining of methanol-fixed HeLa cells using FAK (phospho-Tyr925) antibody (#AP02411PU, Red). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
50 µl / €240.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using FAK antibody. Rabbit Aff - Purified Hu, Ms P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using FAK antibody. Rabbit Aff - Purified Hu, Ms P, WB
50 µl / €230.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using FAK antibody AP02673PU. Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using FAK antibody AP02673PU. Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemistry (IHC) analyzes of FAK antibody (Cat.-No.: AP06600PU-N) in paraffin-embedded human brain tissue. Rabbit Aff - Purified Hu, Ms, Rt P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemistry (IHC) analyzes of FAK antibody (Cat.-No.: AP06601PU-N) in paraffin-embedded human brain tissue. Rabbit Aff - Purified Hu, Ms, Rt P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

FAK1 / PTK2 antibody

Immunohistochemistry: FAK Antibody (EP1831Y) [NB100-79943] - Immunohistochemical staining of paraffin-embedded human tonsils using anti-FAK. Rabbit IgG EP1831Y Hu, Ms, Rt F, ICC/IF, P, WB
0.1 ml / €480.00
  Novus Biologicals Inc.

FAK1 / PTK2 pTyr861 antibody

Western Blot: FAK Phosphospecific Antibody [NB100-81921] - Western blot analysis using FAK (Ab-861) Lane 1 and 2) and FAK (phospho-Tyr861) antibody (#NB100-81921, Lane 3, 4, 5 and 6). Rabbit IgG Aff - Purified Hu, Ms, Rt WB
50 µg / €360.00
  Novus Biologicals Inc.

FAK1 / PTK2 pTyr925 antibody

Western Blot: FAK Phosphospecific Antibody [NB100-81922] - Western blot analysis of extracts using FAK (phospho- Tyr925) antibody Rabbit IgG Aff - Purified Hu, Ms, Rt WB
50 µg / €360.00
  Novus Biologicals Inc.

FAK1 / PTK2 antibody

Western Blot: FAK Antibody [NB600-847] - Western Blot of Native FAK immunoprecipitated from human cells. Rabbit IgG Purified Amph, Can, Hu, Ms, Rt ICC/IF, IP, WB
0.5 ml / €400.00
  Novus Biologicals Inc.

FAK1 / PTK2 pTyr397 antibody

Immunofluorescent: Focal Adhesion Kinase Phosphospecific Antibody (141-9) [NSB625] - Immunofluorescence Staining HeLa cells were fixed prior to immunostaining with the FAK rabbit monoclonal antibody. The signal was detected with an anti-rabbit FITC conjugated secondary antibody. The data show that the antibodydetected phosphoryl Rabbit IgG 141-9 Purified Hu ICC/IF, WB
0.1 ml / €420.00
  Novus Biologicals Inc.

FAK2 / PTK2B pTyr402 antibody

Transactivation of PYK2 Whole cell lysates of serum starved HepG2 tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to PVDF membranes. Immunoblots were probed with mab PYK2-14F6 (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec). lane 1:Co; lane 2: serum; lane 3: EGF; lane 4: H2O2;lane 5: Anisomycin; lane 6: Sorbit; lane 7: Arsen; lane 8:Ceramide. Mouse IgG1 14F6 Purified Hu, Ms, Rt E, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

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