Anti paxillin

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-paxillin is a primary antibody that specifically binds to the paxillin protein. Paxillin is a focal adhesion-associated protein that plays a role in cell signaling and cytoskeletal organization.

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Lab products found in correlation

Close spelling variants of this product

Anti-phosphoTyr118-paxillin Anti-paxillin antibody Rabbit anti-Paxillin Rabbit anti-phospho-paxillin Y118 Anti-p-Paxillin Anti-phospho-Paxillin (Tyr118) Anti-pY118-paxillin Rabbit anti‐Phospho‐Paxillin Anti-phospho-paxillin Paxillin

14 protocols using anti paxillin

Synthesis and Characterization of Phosphoinositide Compounds

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5-PCF₂Am-InsP₅ (CF2) was synthesized as previously described [21] (link). 5-InsP₇ and 5-PCP-InsP₅ (5-PCP) synthesized using similar methods to those previously described [22] (link), [23] (link), [24] (link), [25] . All synthetic compounds were purified by ion-exchange and/or RP-18 chromatography and were fully characterized by ¹H, ³¹ P, and ¹³ C nuclear magnetic resonance spectroscopy.
Anti-IP6K1, anti-myc, anti-coronin 1B, anti-Arp3, anti-cadherin antibodies were from Santa Cruz Biotechnology. Anti-Arp2, anti-p34 antibodies were from Bethyl Laboratories. Anti-flag, anti-vinculin antibodies were from Sigma-Aldrich. Anti-α-actinin, anti-FAK, anti-paxillin, anti-phospho-paxillin, anti-β-actin antibodies were from Cell Signaling Technology. Anti-phospho-FAK (Y397) antibody was from Abcam. Anti-GST antibody was from Proteintech.
HEK293, HEK293T/17 cell lines were from ATCC, HUVECs were from Lonza.

Qi J., Cheng W., Gao Z., Chen Y., Shipton M.L., Furkert D., Chin A.C., Riley A.M., Fiedler D., Potter B.V, & Fu C. (2023). Itraconazole inhibits endothelial cell migration by disrupting inositol pyrophosphate-dependent focal adhesion dynamics and cytoskeletal remodeling. Biomedicine & Pharmacotherapy, 161, 114449.

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Signaling Pathways in Rheumatoid Arthritis

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Due to unique cellular responses observed in HFLS-RA cells (compared to HFLS-OA or HFLS control cells), cell signaling pathways were examined only in HFLS-RA cells. Several signal transduction pathways involved in the transformation of HFLS-RA cells to an aggressive phenotype were examined (16 (link), 50 (link), 59 (link)). Briefly, HFLS cells were incubated with antigens for 15-min, 30-min, 1-h and 4-h, cellular lysates were prepared as above, and the following primary antibodies (all polyclonal rabbit IgGs) were utilized to probe the samples; anti-β-actin antibody (endogenous control) (Novus Biologicals), anti-SAPK/JNK (#9252), anti-phospho-SAPK/JNK (Thr183/Tyr185) (#9251), anti-Erk1/2 (p44/42 MAPK) (#9102), anti-phospho-Erk1/2 (p44/42 MAPK; Thr202/Tyr204) (#9101), anti-Akt (#9272), anti-phospho-Akt (Ser473) (#9271), anti-p38 MAPK (#9212), anti-phospho-p38 MAPK (Thr180/Tyr182) (#4511), anti-Paxillin (#2542), anti-phospho-Paxillin (Tyr118) (#2541), anti-FAK (#3285), anti-phospho-FAK (Tyr397) (#3283) (Cell Signaling Technology, Danvers, MA, USA). HRP-conjugated anti-rabbit IgG (Jackson ImmunoResearch) was used as a secondary antibody. Respective band intensities were measured and reported as outlined above. The maximum and minimum activation of signaling pathways for HFLS lysates was determined based on positive (treatment with LPS) and negative (treatment with media) controls.

Aripova N., Duryee M.J., England B.R., Hunter C.D., Mordeson J.E., Ryan E.M., Daubach E.C., Romberger D.J., Thiele G.M, & Mikuls T.R. (2023). Citrullinated and malondialdehyde-acetaldehyde modified fibrinogen activates macrophages and promotes an aggressive synovial fibroblast phenotype in patients with rheumatoid arthritis. Frontiers in Immunology, 14, 1203548.

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Synthesis and Characterization of Inositol Polyphosphates

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5-PCF₂Am-InsP₅ (CF2) was synthesized as previously described [21 (link)]. 5-InsP₇ and 5-PCP-InsP₅ (5-PCP) synthesized using similar methods to those previously described [22 (link)-25 ]. All synthetic compounds were purified by ion-exchange and/or RP-18 chromatography and were fully characterized by ¹H, ³¹P, and ¹³C nuclear magnetic resonance spectroscopy.
Anti-IP6K1, anti-myc, anti-coronin 1B, anti-Arp3, anti-cadherin antibodies were from Santa Cruz Biotechnology. Anti-Arp2, anti-p34 antibodies were from Bethyl Laboratories. Anti-flag, anti-vinculin antibodies were from Sigma-Aldrich. Anti-α-actinin, anti-FAK, anti-paxillin, anti-phospho-paxillin, anti-β-actin antibodies were from Cell Signaling Technology. Anti-phospho-FAK (Y397) antibody was from Abcam. Anti-GST antibody was from Proteintech.
HEK293, HEK293T/17 cell lines were from ATCC, HUVECs were from Lonza.

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Western Blot Analysis of Focal Adhesion Signaling

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SCs were washed twice with cold PBS and incubated in a lysis buffer (RIPA, BioTeke, Beijing, China) containing 1 mmol/L protease inhibitor PMSF (Sigma, St. Louis, MO, USA) and phosphatase inhibitors (Invitrogen, Carlsbad, CA, USA) on ice, followed by centrifugation at 12,000 g and 4°C for 15 min. Protein concentration was determined using a Micro BCA Protein Assay Kit (Thermo Fisher Scientific, Rockford, USA). Protein samples (15 μg) were loaded onto a 10% polyacrylamide gel and transferred onto PVDF membranes using semidry methodology, as previously described [36 (link)]. Membranes were probed using specific antibodies: anti-FAK (#3285), anti-phospho-FAK (Tyr397; #8556), anti-paxillin (#3283), anti-phospho-paxillin (Tyr118; #2541), anti-Akt (#9272) or anti-phospho-Akt (Ser473; #9271), which were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-β-actin (AP0060), anti-rabbit IgG (BS13278) and anti-mouse IgG (BS30503) were obtained from Bioworld Technology (Louis Park, MN, USA). The proteins were visualized with the ECL-Plus Western blotting reagent in a FluorChem M system (Cell BioSciences, San Leandro, CA, USA). Band density was analyzed by Image J (http://rsb.info.nih.gov/ij/).

Wang D., Gao C.Q., Chen R.Q., Jin C.L., Li H.C., Yan H.C, & Wang X.Q. (2016). Focal adhesion kinase and paxillin promote migration and adhesion to fibronectin by swine skeletal muscle satellite cells. Oncotarget, 7(21), 30845-30854.

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Osimertinib and FAK Inhibitor Signaling

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Osimertinib was purchased from Selleckchem (S7297), and FAK inhibitor 14 and FAK autophosphorylation inhibitor from Abcam (ab144503). The following antibodies were obtained from Cell Signaling Technology: anti-FAK (#3285), anti-Phos-FAK (Tyr397) (#3283), anti-paxillin (#2542), anti-Phos-paxillin (Tyr118) (#69363), anti-thrombospondin-1 (D7E5F) (#37879), anti-AKT3 (#4059), anti-COL5A1(#37304), anti-E-cadherin (#3195), anti-PARP (#9542), anti-caspase-3 (#9662), anti-cleaved PARP (Asp214) (#5625), anti-N-cadherin (D4R1H) (#13116), anti-E-cadherin (24E10) (#3195), anti-vimentin (D21H3) (#5741), anti-rabbit IgG HRP-linked (#7074P2), and anti-mouse IgG HRP-linked (#7076P2).
The anti-CADM1 (PA3-16744) and anti-BICC (PA5-116342) were purchased from Thermo Fisher. The anti-IGFBP7 (ab74169) and anti-PTPRM (ab231607) were purchased from Abcam. The anti-β-actin (A2228) and anti-RAB32 (HPA025731) were purchased from Sigma-Aldrich.

Kosibaty Z., Brustugun O.T., Zwicky Eide I.J., Tsakonas G., Grundberg O., De Petris L., McGowan M., Hydbring P, & Ekman S. (2022). Ras-Related Protein Rab-32 and Thrombospondin 1 Confer Resistance to the EGFR Tyrosine Kinase Inhibitor Osimertinib by Activating Focal Adhesion Kinase in Non-Small Cell Lung Cancer. Cancers, 14(14), 3430.

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Antibody panel for cell signaling study

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Anti PI3KC2α (#22028‐1‐AP, Proteintech), anti GFP (gift from Emilia Turco, University of Turin, Italy), anti α‐tubulin (#2125, Cell Signaling), anti GAPDH (sc‐47724, Santa Cruz Biotechnology), anti Myc‐tag (#2276, Cell Signaling), anti FAK (#71 433, Cell Signaling), anti p‐FAK (tyr397) (#8556, Cell Signaling), anti p‐FAK (tyr925) (#3284, Cell Signaling), anti Paxillin (#2542, Cell Signaling), anti p‐Paxillin (tyr118) (#69 363, Cell Signaling), anti HA‐tag (# 26 183, Thermofisher), anti R‐RAS (#8446, Cell Signaling), anti RASA3 (#PA5‐30445,Invitrogen), anti Rap1A/Rap1B (#4938, Cell Signaling), anti RAS (#3339, Cell Signaling), and anti Vinculin (#V9131, Sigma).

Li H., Prever L., Hsu M.Y., Lo W., Margaria J.P., De Santis M.C., Zanini C., Forni M., Novelli F., Pece S., Di Fiore P.P., Porporato P.E., Martini M., Belabed H., Nazare M., Haucke V., Gulluni F, & Hirsch E. (2022). Phosphoinositide Conversion Inactivates R‐RAS and Drives Metastases in Breast Cancer. Advanced Science, 9(9), 2103249.

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Cell Migration Molecular Mechanisms

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The anti-FNDC4, anti-MYOG and anti-desmin were from Bioss Biotechnology. The anti-GAPDH, anti-Phospho-paxillin (Tyr118), anti-paxillin, anti-Phospho-FAK (Tyr925), anti-FAK, anti-Vinculin and anti-ITGβ1 were purchased from Cell Signaling. The anti-6× his and anti-IgG were from Santa Cruze Biotechnology. The secondary antibodies were HRP-labeled and FITC-labeled were from Beijing Biosynthesis Biotechnology Co., Ltd. PF562271 was from Selleck and used concentration for 10 nM/ml to treat cells.

Wang Z., Wang Z., Pang Y., Tong H., Yan Y., Li S, & Li S. (2020). Fibronectin type III domain-containing 4 promotes the migration and differentiation of bovine skeletal muscle-derived satellite cells via focal adhesion kinase. Cell Adhesion & Migration, 14(1), 153-164.

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Antibodies and Reagents for Cell Signaling

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Anti-SCGN antibody was from AbFrontier. Anti-FAK, anti-paxillin, anti-phospho-paxillin (Tyr¹¹⁸), anti-ERK1/2, anti-phospho-ERK1/2 (Thr²⁰²/Tyr²⁰⁴), anti-Akt and anti-phospho-Akt (Ser⁴⁷³) antibodies were from Cell Signaling Technology. Anti-α-tubulin antibody, anti-β-actin antibody and normal rabbit IgG were from Santa Cruz Biotechnology. Anti-phospho-FAK (Tyr³⁹⁷) and anti-SCGN antibodies used in immunoprecipitation were from Abcam. anti-paxillin antibody used in confocal microscopy was from Millipore Corporation. Anti-E-cadherin (epithelial cadherin) and anti-N-cadherin (neural cadherin) antibodies were from BD Biosciences. Horseradish peroxidase-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were from Bio-Rad Laboratories. Rhodamine–phalloidin, Alexa Fluor® 488- or Alexa Fluor® 568-conjugated goat anti-rabbit IgG and Alexa Fluor® 488-conjugated goat anti-mouse IgG were from Invitrogen. Latrunculin B was from Calbiochem. Cytochalasin D, ionomycin and DMSO from Sigma–Aldrich. Penicillin G, streptomycin, FBS and trypsin were from Gibco Life Technologies. DMEM (Dulbecco's modified Eagle's medium) and 45% D-glucose were from WelGENE. SMARTpool siRNA and DharmaFECT1 transfection reagent were from Dharmacon. Insulin ELISA kit was from ALPCO. BCA protein assay was from Thermo Scientific. Protein G–Sepharose beads and silver staining kit were from GE Healthcare.

Yang S.Y., Lee J.J., Lee J.H., Lee K., Oh S.H., Lim Y.M., Lee M.S, & Lee K.J. (2016). Secretagogin affects insulin secretion in pancreatic β-cells by regulating actin dynamics and focal adhesion. Biochemical Journal, 473(12), 1791-1803.

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Integrin and Cytoskeleton Dynamics in HA-Stimulation

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Primary antibodies for anti-CD44 (R&D systems, UK), anti-cortactin (Upstate Biotechnology, NY, USA), anti-paxillin, anti-phosphoTyr¹¹⁸-paxillin were obtained from Cell Signaling Technology (Beverly, MA), while anti-α5β1-integrin, anti-α2β1-integrin, anti-β1-integrin, activated-β1-integrin conformations (B44 and HUTS-4), cellular Fibronectin and phosphoTyr⁴²¹-cortactin were purchased from Millipore (Watford, UK). HRP-conjugated secondary antibodies were obtained from Amersham, UK and Hyaluronan (MW220 kDa) from Lifecore Biomedical (MN, USA). Cells were stimulated with 100 μg/ml HA for indicated times. All other reagents were purchased from Sigma (Poole, UK) unless otherwise stated.

McFarlane S., McFarlane C., Montgomery N., Hill A, & Waugh D.J. (2015). CD44-mediated activation of α5β1-integrin, cortactin and paxillin signaling underpins adhesion of basal-like breast cancer cells to endothelium and Fibronectin-enriched matrices. Oncotarget, 6(34), 36762-36773.

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Analysis of Cytoskeletal Regulators

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Anti-phospho-cofilin (Ser-3), anti-cofilin, anti-phospho-MLC (Thr-18/Ser-19), anti-MLC, anti-phopsho-LIMK1 (Thr-508)/LIMK2 (Thr-505), anti-LIMK1, anti-phospho-Src family (Tyr-416), anti-phospho-Src (Tyr-527), anti-Src, anti-phospho-FAK (Tyr-397), anti-FAK, anti-α-actinin, anti-paxillin, anti-talin-1, and anti-tensin-2 antibodies were purchased from Cell Signaling Technology. Anti-PCTK3 and anti-vinculin antibodies were from Santa Cruz Biotechnology. Anti-FLAG (M2) antibody was form Sigma-Aldrich. Anti-Strep antibody was from Qiagen. Anti-Halo antibody was from Promega. Anti-GAPDH antibody was from Wako Pure Chemical Industries. Anti-RhoA and Rac1 antibodies and Alexa-555 conjugated-phalloidin were from Cytoskeleton. Anti-Myc antibody was from Enzo Life Sciences.

Matsuda S., Kawamoto K., Miyamoto K., Tsuji A, & Yuasa K. (2017). PCTK3/CDK18 regulates cell migration and adhesion by negatively modulating FAK activity. Scientific Reports, 7, 45545.

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