Vinculin v9264

Manufactured by Merck Group

Sourced in United States

Vinculin (V9264) is a protein that plays a crucial role in cell-cell and cell-matrix adhesion. It is a widely used marker for focal adhesions and adherens junctions in cell biology research.

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Lab products found in correlation

Polyclonal erk m5670, by Merck Group (3 mentions) Image lab software, by Bio-Rad (2 mentions) α tubulin 2144, by Cell Signaling Technology (2 mentions) Perk sc 7383 and sc 16982 r, by Santa Cruz Biotechnology (2 mentions) Etv5 wh0002119m2, by Merck Group (2 mentions) Gfp sc 9996, by Santa Cruz Biotechnology (2 mentions) Ha c29f4, by Cell Signaling Technology (2 mentions) Oligomycin, by Merck Group (1 mentions) Thioacetamide taa, by Merck Group (1 mentions) Cd83 pe, by BD (1 mentions) F4 80 bm8, by Thermo Fisher Scientific (1 mentions) Hla dr apc h7, by BD (1 mentions) Cd1a fitc, by BD (1 mentions) Cd86 bv421, by BD (1 mentions) Cd40 apc, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Cd14 v500, by BD (1 mentions) Flag m2 monoclonal, by Merck Group (1 mentions) P mek1 2, by Santa Cruz Biotechnology (1 mentions) Pshp2 y542, by Abcam (1 mentions)

9 protocols using vinculin v9264

Molecular Profiling of Cell Subsets

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Thioacetamide (TAA), Oligomycin, DMSO, and CCCP were purchased from Sigma‐Aldrich. Vinculin #V9264, STAT1 #SAB4300326, and CYP2E1 HPA009128 were obtained from Sigma‐Aldrich, while p100/p52 #06‐413 from Millipore (Billerica, MA, USA) and p105/p50 #D4P4D from Cell Signaling (Danvers, MA, USA). PRC antibody was described previously.⁵ Anti‐mouse: CD45 (30‐F11), CD11c (HL3), Ly6c (AL‐21), Ly6g (1A8), MHCII I‐A/I‐E (M5/114.15.2), CD19 (ID3), CD8 (53‐6.7), CD4 (RM4‐5), TCRβ (H57.597), CD11b (M1/70) were purchased from BD Biosciences and F4/80 (BM8) from eBioscience. Anit‐human: CD14‐v500, CD1a‐FITC, HLA‐DR‐APC‐H7, CD86‐BV421, CD83‐PE were acquired from BD Biosciences and CD40‐APC from eBiosciences.

Buler M., Naessens T., Mattsson J., Morias Y., Söderberg M., Robbins P., Kärrberg L., Svensson T.S., Thulin P., Glinghammar B., Scarpulla R.C, & Andersson U. (2020). The regulatory role of PGC1α‐related coactivator in response to drug‐induced liver injury. FASEB BioAdvances, 2(8), 453-463.

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Immunoblotting Antibody Validation Protocol

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Rabbit polyclonal antibodies against Src (#2109, 1:5000), phosphorylated (p)Src (#2101, 1:1000), pAKT (#9271, 1:1000), AKT (#9272, 1:1000), ERK (#9102, 1:1000), SYK (#2712, 1:1000), FAK (#3285, 1:1000), pTyr (P-Tyr-1000) (#8954, 1:2000), cleaved caspase-3 (#9664, 1:1000), cleaved PARP (#9541, 1:1000), PARP (#9542, 1:1000), cleaved caspase-9 (#9661, 1:1000), caspase-9 (#9508, 1:1000), and HA (#3724, 1:5000) were obtained from Cell Signaling Technologies. Polyclonal IgG (sc-2027), pERK (sc-7383, 1:500), CBL (sc-170, 1:500), SHP2 (sc-280, 1:1000), pMEK1/2 (sc-81503, 1:500), and MEK-1 (sc-6250, 1:500) were obtained from Santa Cruz Biotechnology. p-SHP2(Y542) (ab62322, 1:20,000) was obtained from Abcam. Monoclonal antibodies against Pan-Ras (OP40, 1:500), HA (12CA5, 1:500), and pTyr (4G10) (05–321, 1:1000) were obtained from Boehringer Ingelheim and Millipore, respectively. Monoclonal FLAG-M2 (F1804, 1:2000) and Vinculin (V9264, 1:2000) were obtained from Sigma.

Kano Y., Gebregiworgis T., Marshall C.B., Radulovich N., Poon B.P., St-Germain J., Cook J.D., Valencia-Sama I., Grant B.M., Herrera S.G., Miao J., Raught B., Irwin M.S., Lee J.E., Yeh J.J., Zhang Z.Y., Tsao M.S., Ikura M, & Ohh M. (2019). Tyrosyl phosphorylation of KRAS stalls GTPase cycle via alteration of switch I and II conformation. Nature Communications, 10, 224.

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Recombinant Protein Characterization Protocol

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Purified recombinant GST-H-Ras(WT) was obtained from Jena Bioscience. Active recombinant human His-c-Src kinase and GST-SHP2 were obtained from GenWay Biotechnology. Rabbit polyclonal antibodies against Src, phosphorylated (p)Src(Y416), pAKT, AKT, glutathione S-transferase (GST), pSHP2(Y542), SHP2 and HA (polyclonal) were obtained from Cell Signaling Technologies. Polyclonal IgG (sc-2027), pERK (sc-7383), N-Ras (sc-519), and c-Raf (sc-7267) were obtained from Santa Cruz Biotechnology. p-c-Raf(Y340) (#44506G) was obtained from Invitrogen. GFAP and Ki67 were obtained from Dako. Monoclonal antibodies against HA (12CA5) and pTyr (4G10) (05-321) were obtained from Boehringer Ingelheim and Millipore, respectively. Monoclonal FLAG-M2 (F1804), β-actin (A5316) (1:10,000), vinculin (V9264) (1:10,000) and polyclonal ERK (M5670) antibodies (1:5,000) were obtained from Sigma. All antibodies were utilized at a 1:1,000 dilution unless otherwise specified.

Bunda S., Burrell K., Heir P., Zeng L., Alamsahebpour A., Kano Y., Raught B., Zhang Z.Y., Zadeh G, & Ohh M. (2015). Inhibition of SHP2-mediated dephosphorylation of Ras suppresses oncogenesis. Nature Communications, 6, 8859.

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Antibody Panel for Protein Detection

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Antibodies used in the study are β-actin (AC-15), flag epitope (F1804), myc (M4439), PABP (P6246) and Vinculin (V9264) from Sigma-Aldrich; TIA-1 (sc-1751) and p-c-Jun (Ser63/73, sc-16312) from Santa Cruz Biotechnology; RFP antibody (GTX59862) and Vinexin (GTX115362) from GeneTex; GFP (#29779) from AnaSpec; c-Jun (#9165) from Cell Signaling Technology and AlexaFluor 488, 594 or 647-conjugated secondary antibodies from Invitrogen. The CPEB4 monoclonal antibody was raised using the N-terminal 458 amino acids (a.a.) of rat CPEB4 (rCPEB4) produced in Escherichia coli. The CPEB2 antibody has been described elsewhere [10] (link). All chemicals were obtained from Sigma-Aldrich.

Chang Y.W, & Huang Y.S. (2014). Arsenite-Activated JNK Signaling Enhances CPEB4-Vinexin Interaction to Facilitate Stress Granule Assembly and Cell Survival. PLoS ONE, 9(9), e107961.

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Antibody Panel for Ras Signaling

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A 12-mer phospho-peptide comprising the KRAS sequence flanking pTyr64 (TAGQEEpYSAMRD) was used to generate anti-pRAS Y64 rabbit polyclonal antibody (1 : 1000; Bethyl Laboratory, Inc.). Pan-Ras (OP40, 1 : 500), mAbs against HA (12CA5, 1 : 500), KRAS (OP24, 1 : 200, 1 : 500), and pTyr (4G10) (05-321, 1 : 1000) were obtained from Boehringer Ingelheim and Millipore, respectively. Monoclonal FLAG-M2 (F1804, 1 : 2000), β-actin (A5316, 1 : 2000) and Vinculin (V9264, 1 : 2000) antibodies were obtained from Sigma. Rabbit polyclonal antibodies against Src (#2109, 1 : 5000), pERK (#9101, 1 : 1000), ERK (#9102, 1 : 1000), pTyr (P-Tyr-1000) (#8954, 1 : 2000), Ras (3965 S, 1 : 1000), and HA (#3724, 1 : 5000) were obtained from Cell Signaling Technologies. HA.11 (#16B12, 1 : 1000) was obtained from Covance. Polyclonal IgG (sc-2027), HRAS (sc-520, 1 : 1000), NRAS (sc-519, 1 : 1000), and SHP2 (sc-280, 1 : 1000) were obtained from Santa Cruz Biotechnology.

Gebregiworgis T., Kano Y., St-Germain J., Radulovich N., Udaskin M.L., Mentes A., Huang R., Poon B.P., He W., Valencia-Sama I., Robinson C.M., Huestis M., Miao J., Yeh J.J., Zhang Z.Y., Irwin M.S., Lee J.E., Tsao M.S., Raught B., Marshall C.B., Ohh M, & Ikura M. (2021). The Q61H mutation decouples KRAS from upstream regulation and renders cancer cells resistant to SHP2 inhibitors. Nature Communications, 12, 6274.

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Immune Cell Lysis and Immunoblotting

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5x10⁶ B-cells were lysed with RIPA Buffer (20 mM HEPES at pH 7.5, 300 mM NaCl, 5 mM EDTA, 10% Glycerol, 1% Triton X-100, supplemented with protease inhibitors (Roche) and phosphatase inhibitors (0.4 mM Orthovanadate, 10 mM NaF) and briefly sonicated. Cleared lysates were electrophoresed and immunoblotted with the indicated primary antibodies: c-Myc Y69 (ab32072) from Abcam, Vinculin (V9264) from Sigma. Chemioluminescent detection, after incubation of the membranes with appropriate secondary antibodies, was done through a CCD camera using the ChemiDoc System (Bio-Rad). Quantification of protein levels was done using the Image Lab Software (Bio-Rad, version 4.0).

Tesi A., de Pretis S., Furlan M., Filipuzzi M., Morelli M.J., Andronache A., Doni M., Verrecchia A., Pelizzola M., Amati B, & Sabò A. (2019). An early Myc‐dependent transcriptional program orchestrates cell growth during B‐cell activation. EMBO Reports, 20(9), e47987.

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Protein Expression Analysis by Immunoblotting

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5×10⁶ to 10×10⁶ cells were lysed with RIPA Buffer (50 mM Tris-HCl pH 8, 150 mM NaCl, 5 mM EDTA, 0.5% NP-40, 0.1% SDS, supplemented with protease inhibitors (Mini, Roche) and phosphatase inhibitors 0.2 mM Ortovanadate, 5 mM NaF) and sonicated. Cleared lysates were electrophoresed and immunoblotted with the indicated primary antibodies: p53 (NCL-p53-CM5p) from Novocastra laboratories, phospho-histone H2A.X (Ser139) clone JBW301 from Millipore and Vinculin (V9264) from Sigma. After incubation of the membranes with appropriate secondary antibodies, imaging was performed using an enhanced chemiluminescence (ECL) detection kit (Bio-Rad), followed by analysis with ChemiDoc XRS+ imaging system and Image Lab software (Bio-Rad).

Tonelli C., Morelli M.J., Bianchi S., Rotta L., Capra T., Sabò A., Campaner S, & Amati B. (2015). Genome-wide analysis of p53 transcriptional programs in B cells upon exposure to genotoxic stress in vivo. Oncotarget, 6(28), 24611-24626.

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Antibody Source and Dilution Protocol

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The following antibodies were obtained from Cell Signaling Technologies: HA (C29F4) (1:6000), Lamin(A/C) (2032), pERK (4370), ubiquitin (3933), PJA2 (40180), and β-TrCP (D13F10), β-actin (8H10D10) (1:20,000), myc tag (71D10), and α-tubulin (2144) (1:5000). pERK (sc-7383 and sc-16982-R), PJA1 (sc-517068), and GFP (sc-9996) (1:6000) were obtained from Santa Cruz Biotechnology. Ki67 was obtained from Dako. Phospho Ser/Thr (ab17464), T7 tag (ab9138), capicua (ab123822), ETV1 (ab81086), Renilla (ab187338), and myc protein (ab3207) were obtained from Abcam. The following antibodies were purchased from Millipore T7 tag (69522), capicua (ABN446) and capicua (MABN449). FLAG-M2 (F1804), β-actin (A5316) (1:10,000), vinculin (V9264) (1:30,000), ETV5 (WH0002119M2), and polyclonal ERK (M5670) antibodies (1:5000) were obtained from Sigma. PJA1 (MBS153701) was purchased from MyBioSource.com. All antibodies were utilized at a 1:1,000 dilution unless otherwise specified.

Bunda S., Heir P., Metcalf J., Li A.S., Agnihotri S., Pusch S., Yasin M., Li M., Burrell K., Mansouri S., Singh O., Wilson M., Alamsahebpour A., Nejad R., Choi B., Kim D., von Deimling A., Zadeh G, & Aldape K. (2019). CIC protein instability contributes to tumorigenesis in glioblastoma. Nature Communications, 10, 661.

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Antibody Characterization for Cell Signaling

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The following antibodies were obtained from Cell Signaling Technology: HA (C29F4) (1:6,000), c-Src (2108), pSrc(416) (2101), Lamin(A/C) (2032), pERK (4370), β-actin (8H10D10) (1:20,000), and α-tubulin (2144) (1:5,000). pERK (sc-7383 and sc-16982-R) and GFP (sc-9996) (1:6,000) were obtained from Santa Cruz Biotechnology. Phospho Ser/Thr (ab17464), capicua (ab123822), and ETV1 (ab81086) were obtained from Abcam. The following antibodies were purchased from Millipore: capicua (ABN446), capicua (MABN449), EGFR, and pTYR. FLAG-M2 (F1804), β-actin (A5316) (1:10,000), vinculin (V9264) (1:30,000), ETV5 (WH0002119M2), and polyclonal ERK (M5670) antibodies (1:5,000) were obtained from Sigma. All antibodies were utilized at a 1:1,000 dilution unless otherwise specified.

Bunda S., Heir P., Li A.S., Mamatjan Y., Zadeh G, & Aldape K. (2020). c-Src Phosphorylates and Inhibits the Function of the CIC Tumor Suppressor Protein. Molecular cancer research : MCR, 18(5), 774-786.

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