We used the following primary antibodies: anti-FN (1:1,000, Millipore, Burlington MA USA), anti-α-SMA-Cy3 conjugated (1:500, Sigma-Aldrich, St. Louis, MO USA), anti-paxillin (1:300, BD Transduction, Spain), anti-keratin 6 (1:500, Covance, Spain), anti p-Smad2/3 (1:300, Thermo Fisher, Spain), anti-α5 integrin (1:100, Cell Signaling Technology, The Netherlands) and anti-Ki67 (1:200, Abcam, Cambridge, UK). For flow cytometry, we used anti-α5 integrin (1:200 BD Pharmingen, Spain) and anti β3 integrin (1:200 BD Bioscience, Spain). Secondary antibodies: anti-rabbit Alexa Fluor 488 (1:400 Thermo Fisher, Spain) and anti-mouse Alexa Fluor 647 (1:500, Invitrogen). To stain the cytoskeleton, we used Rhodamine Phalloidin (1:500 Thermo Fisher). To stain nuclei, we used either DAPI (1:10,000, Thermo Fisher) or Hoechst (1:10,000, Thermo Fisher).
Anti integrin α5
Manufactured by Cell Signaling Technology
Sourced in Spain, Germany
Anti-Integrin α5 is a laboratory reagent that recognizes the α5 subunit of the integrin receptor. Integrins are transmembrane receptors that mediate cell-cell and cell-extracellular matrix interactions. The α5 subunit is a component of the α5β1 integrin, which binds to the extracellular matrix protein fibronectin.
Automatically generated - may contain errors
Lab products found in correlation
Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti fak, by Cell Signaling Technology (2 mentions)
Anti integrin β1, by Cell Signaling Technology (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Anti paxillin, by BD (1 mentions)
Anti fn, by Merck Group (1 mentions)
Anti p smad2 3, by Thermo Fisher Scientific (1 mentions)
Anti integrin α5, by BD (1 mentions)
Anti ki67, by Abcam (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Anti eea1, by Cell Signaling Technology (1 mentions)
Anti integrin αv, by Abcam (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Anti rab5, by Cell Signaling Technology (1 mentions)
Anti lamp1, by Cell Signaling Technology (1 mentions)
Anti rab9, by Cell Signaling Technology (1 mentions)
6 protocols using anti integrin α5
1
Immunohistochemistry and Flow Cytometry Protocols
2
Integrin and Vesicle Trafficking Imaging Protocol
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The following primary antibodies were used for immunoblotting: anti-α5-integrin (Merck Millipore), anti-αv-integrin (Abcam, Cambridge, UK), anti-β1-integrin (BD Transduction Laboratories, San Jose, CA, USA), anti-β3-integrin (Merck Millipore), anti-β5-integrin (Abcam), anti-pERK (Cell Signaling Technology, Danvers, MA, USA), anti-ERK (Cell Signaling Technology), anti-LAMP1 (Cell Signaling Technology) and anti-β-actin (Sigma-Aldrich). The following primary antibodies were used for immunofluorescence staining: anti-α5-integrin (Cell Signaling Technology), anti-αv-integrin (Cell Signaling Technology), anti-LAMP1 (Cell Signaling Technology), anti-EEA1 (Cell Signaling Technology), anti-Rab5 (Cell Signaling Technology), anti-Rab7 (Cell Signaling Technology), anti-Rab9 (Cell Signaling Technology) and anti-γ-H2AX (Merck Millipore).
3
Immunoblotting Analysis of Cell Signaling
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Tissue samples were snap frozen and stored at − 80 °C or lysed in RIPA-buffer containing protease inhibitors and analyzed by immunoblotting. Primary antibodies used in this study were rabbit anti-zyxin (B71, provided by M. Beckerle, Huntsman Cancer Institute), anti-α-tubulin (#2144, Cell Signaling), anti-α-actinin (#A7811, Sigma-Aldrich), anti-FAK (#3285, Cell Signaling), anti-pFAK (Y397) (#44-624G, Thermo Fisher Scientific), anti-AKT (#9272, Cell Signaling), anti-pAKT (Ser473) (#9271, Cell Signaling), anti-Integrin α5 (#4705, Cell Signaling), anti-Integrin β1 (#9699, Cell Signaling), anti-Col1α2 (#sc-166865, Santa Cruz Biotechnology) and anti-LOX (sc-373995, Santa Cruz Biotechnology). HRP-conjugated anti-mouse (#4416, Sigma-Aldrich) and anti-rabbit (#6154, Sigma-Aldrich) antibodies were used as secondary antibodies and chemiluminescence was detected with the ImageQuant LAS mini system (GE Healthcare). Densitometric analysis was done with ImageJ.
4
Western Blot Analysis of Cell Signaling
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Cells were washed with PBS and lysed with SDS lysis buffer. Frozen tissues were lysed using T-PER Tissue Protein Extraction Reagent (Thermo Fisher Scientific). Protein concentrations were determined using the Bradford assay (Bio-Rad). Equal amounts of protein from each sample were resolved by SDS-PAGE and transferred onto PVDF membranes (Millipore). Immunoblots were blocked by incubation in 5% skim milk in TBS-T (0.1% Tween 20) for 1 h at 25 °C. Membranes were incubated with the following primary antibodies: anti-TINAGL1 (1:1000; 12077-1-AP, Proteintech), anti-pFAK (1:1000; #3283; Cell Signaling Technology), anti-FAK (1:1000; #3285; Cell Signaling Technology), β-actin (1:10,000; sc-47778, Santa Cruz Biotechnology), anti-Twist (1:1000; ab49254, Abcam), anti-E-cadherin (1:500; #13-1700; Invitrogen), anti-N-cadherin (1:1000, ab18203, Abcam), anti-integrin β1 (1:1000; ab30394, Abcam), anti-integrin αv (1:1000; #4711, Cell Signaling Technology), anti-integrin α5 (1:1000; #4705, Cell Signaling Technology), and anti-GAPDH antibodies (1:20,000; Abc-1001, AbClon), followed by their corresponding HRP-conjugated secondary antibodies, anti-mouse (1:5000: #115-035-003, Jackson ImmunoResearch Labs) and anti-rabbit antibodies (1:4000; #Abc-5003, AbClon). Proteins were detected using AbSignal (Abclone).
5
Western Blot Analysis of EDI3 Protein
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Whole cell lysates were collected in RIPA lysis buffer (1% (v/v) NP-40, 150 mM NaCl, 20 mM Tris/HCl (pH 7.4), 1 mM EDTA, 10 mM NaF, 1 mM ZnCl2, 1 mM MgCl2, 1 mM Na3VO4, 10% (v/v) Glycerol) with protease and phosphatase inhibitors (Sigma) and protein content was quantified using the BCA Protein Assay kit (Thermo Scientific). Protein samples were separated on 8% SDS polyacrylamide gels using the mini-PROTEAN Tetra Electrophoresis System (Bio-Rad) and transferred to PVDF membranes (Perkin-Elmer). Primary antibody incubation was performed overnight at 4°C with the following antibodies used at a dilution of 1:1,000 – custom-made mouse monoclonal anti-EDI3 (AMS Bio), anti-β-Actin (Sigma), anti-α-Tubulin, anti-Calnexin, anti-Integrin α5, anti-Integrin β1 (all from Cell Signaling). HRP-conjugated secondary antibodies (anti-rabbit IgG and anti-mouse IgG) were purchased from Cell Signaling and used at a dilution of 1:1,000 for 2 h at room temperature. All images were taken on a Vilber Fusion Fx7 imager (Vilber Lourmat) using chemiluminescence (Perkin-Elmer) for detection and bands were quantified using ImageJ software (National Institute of Health).
6
Antibody Procurement for B19V Research
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The following primary antibodies were purchased: mouse monoclonal antibody of anti-B19V capsid (#MAB8292, clone 521-5D) and mouse anti–β-actin (#A5441) from MilliporeSigma; mouse monoclonal antibody against B19V conformational epitope in VP2 (MAB 860-55D) from MIKROGEN (Neuried, Germany); goat anti-AXL antibody (#AF154) from R&D Systems Minneapolis, MN); rabbit anti-AXL (#8661S, clone C89E7), anti-Ku80 (#2180), and anti-integrin α5 (#98204) from Cell Signaling Technology; and mouse anti-Flag (#200-301-B13) and rabbit anti-BrdU (#600-401-C29) from Rockland, Limerick, PA. Anti-B19V VP1u antibody was in-house prepared.
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