Dimethyl sulfoxide (DMSO), 1,4-NQ (98% purity determined by gas chromatography) and anti-GAPDH antibody were from Wako Pure Chemical Industries (Osaka, Japan), Tokyo Chemical Industries (Tokyo, Japan) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively. Biotin-PEAC5-maleimide (BPM) and Na2S4 were from Dojindo Laboratories (Kumamoto, Japan). Dynabeads M-280-sheep anti-rabbit immunoglobulin G (IgG) was from Invitrogen (Carlsbad, CA, USA). Anti-Akt, anti-CREB, anti-phosphorylated Akt (Ser473), anti-phosphorylated CREB (Ser133), horseradish peroxidase (HRP)-conjugated anti-biotin antibodies, anti-rabbit antibodies and anti-mouse IgG secondary antibodies were from Cell Signaling Technology (Beverly, MA, USA). Escherichia coli BL21 cells and trypsin were from Promega Co. (Madison, WI, USA). Glutathione 4B Sepharose was from GE Healthcare (Chicago, IL, USA). All other reagents were of the highest purity available.
Anti gapdh antibody
Manufactured by Fujifilm
Sourced in Japan
The Anti-GAPDH antibody is a reagent used in laboratory research applications. It is designed to detect the presence and quantify the levels of the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein, which is a commonly used reference or 'housekeeping' protein in various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Ecl detection system, by GE Healthcare (2 mentions)
Complete mini protease inhibitor cocktail, by Roche (2 mentions)
Na2s4, by Dojindo Laboratories (1 mentions)
Biotin peac5 maleimide bpm, by Dojindo Laboratories (1 mentions)
Escherichia coli bl21 cells, by Promega (1 mentions)
Glutathione sepharose 4b, by GE Healthcare (1 mentions)
Anti rabbit antibodies, by Cell Signaling Technology (1 mentions)
Anti mouse igg secondary antibody, by Cell Signaling Technology (1 mentions)
Anti flag, by Merck Group (1 mentions)
Western lightning plus ecl, by Thermo Fisher Scientific (1 mentions)
Anti strep antibody, by Qiagen (1 mentions)
Anti vinculin antibody, by Santa Cruz Biotechnology (1 mentions)
Anti α actinin, by Cell Signaling Technology (1 mentions)
Anti phospho fak tyr 397 anti fak, by Cell Signaling Technology (1 mentions)
Anti talin 1, by Cell Signaling Technology (1 mentions)
Anti halo antibody, by Promega (1 mentions)
Anti paxillin, by Cell Signaling Technology (1 mentions)
Anti flag m2 antibody, by Merck Group (1 mentions)
Ecl film, by GE Healthcare (1 mentions)
Ecl prime blocking agent, by GE Healthcare (1 mentions)
8 protocols using anti gapdh antibody
1
Investigating Protein Interactions Using Biotin-Maleimide
2
Western Blotting Protocol for CORO1C and GAPDH
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The Western blotting procedure has been described previously [24 (link),25 (link),26 (link)]. The anti-CORO1C antibody (Proteintech Group, Inc., Rosemont, IL, USA) was diluted 1:2500, and the anti-GAPDH antibody (Wako, Osaka, Japan), used as the internal control, was diluted 1:1600. The antibodies used are listed in Table S1 .
3
Immunoblotting Protein Detection Protocol
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Immunoblotting was performed as described previously (Kitamura et al., 2017 (link)), using anti-FLAG (1 μg/mL; Merck, Darmstadt, Germany) or anti-GAPDH antibody (1:2000 dilution; FUJIFILM Wako Pure Chemical, Osaka, Japan) as the primary antibody and anti-mouse IgG, HRP-linked F(ab′)2 fragment (1:7500 dilution; GE Healthcare Life Sciences, Little Chalfont, UK) as the secondary antibody. Chemiluminescence was performed using Western Lightning Plus-ECL (Thermo Fischer Scientific, Waltham, MA).
4
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.
5
Western Blot Analysis of Protein Quantification
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Cells were lysed using buffer consisting of 1% Triton X-100, 0.1% sodium dodecyl sulfate (SDS), 150 mM NaCl, 50 mM Tris-HCl, 5 mM Ethylenediaminetetraacetic acid (EDTA), and proteinase inhibitor cocktail (Roche, 04-693-124-001). The protein concentration was determined by DC Protein Assay. Equal amounts of lysates were separated by SDS-PAGE and transferred to PVDF membranes (GE Healthcare, 10600100) using a semidry blotting system in transfer buffer (25 mM Tris base, 190 mM glycine, 20% methanol). The membranes were subsequently blocked with 2% ECL Prime blocking agent (GE Healthcare, RPN418V) and incubated with primary antibodies, followed by corresponding ECL peroxidase labelled secondary antibodies (1:10,000, GE Healthcare). Immunoreactivity was detected by the ECL reaction (GE Healthcare, RPN2235). The blots were exposed to ECL films (GE Healthcare) before they were visualized by scanning densitometry. After detection of the protein of interest, to control for equal loading, blots were submerged in stripping buffer (62.5 mM Tris-HCl pH 6.7, 100 mM 2-mercaptoethanol, and 2% SDS) for 30 min at 50 °C before blocking, and then reprobed with mouse monoclonal anti-GAPDH antibody (1:1000, Wako, 014-25524). Quantitative measurement of bands were obtained by Image J software and expressed as a ratio of the amount of protein of interest relative to GAPDH.
6
Analysis of β-catenin and MAPK Signaling
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Subcloned cells were lysed in TNE buffer with cOmplete Mini Protease Inhibitor Cocktail (Roche), and 10 μg of the protein sample was separated using 10% SDS‐polyacrylamide gel. Antibodies against active (dephosphorylated) β‐catenin (8E7; Millipore, 05–665), total β‐catenin (#9562), phospho‐p44/42 Erk1/2 (#9101), ERK (#9102), and p53 (1C12; Cell Signaling, #2524) were used as primary antibodies. Anti‐GAPDH antibody (Wako, 016‐25523) was used as a control. The ECL detection system (GE Healthcare) was used to detect the signals.
7
Organoid Protein Expression Analysis
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Organoid cells were lysed in TNE buffer with complete Mini protease inhibitor cocktail (Roche), and 10 μg of the protein sample was separated using a 10%–15% SDS-polyacrylamide gel. Antibodies against phosphorylated-p44/42 Erk1/2 (Cell Signaling Technology, #9101), p53 (1C12; Cell Signaling Technology, #2524), HMGA2 (Cell Signaling Technology, #8197S), and HMGA1 (Abcam, #ab129153) were used as primary antibodies. Anti-GAPDH antibody (FUJIFILM Wako, #016–25523, RRID: AB_2814991) was used as the control. An ECL detection system (GE Healthcare) was used to detect signals.
8
Western Blot Analysis of LRP1, Tau, and GAPDH
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Fractionated brain homogenates were dissolved in lithium dodecyl sulfate sample buffer (Novex) containing 2% β-mercaptothanol. Proteins were separated in Tris-Glycine gel and transferred to poly vinylidene difluoride membranes and incubated with an anti-LRP1 antibody 11H4 (a kind gift from Dr. Ayae Kinoshita, Kyoto University, Kyoto, Japan), an anti-actin antibody (A2066-100uL; Sigma-Aldrich), an anti-human tau HJ8.5 antibody (a kind gift from Dr. David Holtzman, Washington University, St. Louis, MO), or an anti-GAPDH antibody (016-25523; FUJIFILM Wako Pure Chemical Corporation), followed by species-specific HRP-conjugated secondary antibodies. Bands were visualized using ImmunoStar (FUJIFILM Wako Pure Chemical Corporation), and signals were detected using ImageQuant LAS-4000 (GE healthcare). Bands were quantified using ImageQuant TL (GE healthcare).
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