Tissue protein and cells were lysed with TNTE buffer which contained protease inhibitors and ten micrograms of protein were loaded onto 12% sodium dodecyl sulphate (SDS)-polyacrylamide gels and the detailed procedures have been described previously [46 (link), 47 (link)]. The protein samples were analyzed using primary antibodies include polyclonal rabbit anti-PTBP1 (1:3,000; provided by Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College) and mouse anti-GAPDH (1:3000; Abmart) antibodies.
Mouse anti gapdh
Manufactured by Abmart
Sourced in China
Mouse anti-GAPDH is a primary antibody that specifically binds to the GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase) protein, which is a widely used loading control and housekeeping gene in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (2 mentions)
Tissue protein extraction reagent, by CWBIO (1 mentions)
Mouse anti th, by Merck Group (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Bca protein assay kit, by CWBIO (1 mentions)
Radioimmunoprecipitation assay (ripa), by CWBIO (1 mentions)
Mouse anti actin, by Merck Group (1 mentions)
Rabbit anti vimentin, by Proteintech (1 mentions)
Rabbit anti β catenin, by Proteintech (1 mentions)
Rabbit anti cpt1a, by Proteintech (1 mentions)
Rabbit anti c6orf15, by Proteintech (1 mentions)
Rabbit anti laminb1, by Proteintech (1 mentions)
Ecl assay kit, by Thermo Fisher Scientific (1 mentions)
4 protocols using mouse anti gapdh
1
Immunoblotting of PTBP1 and GAPDH
2
Zebrafish Brain Tyrosine Hydroxylase Quantification
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The level of tyrosine hydroxylase protein in the zebrafish brain was detected by western blot analysis. After exposure for 7 days, the brains of adult zebrafish were harvested and immediately lysed in a tissue protein extraction reagent (CWBIO, Beijing, China) and 5 μL PMSF (Sigma-Aldrich). The protein concentrations were quantified using the BCA Protein Assay Kit (CWBIO). The proteins were subjected to SDS-PAGE and transferred onto a nitrocellulose membrane blocked with 5% non-fat dry milk in Tris-buffered saline with 0.05% Tween-20. The membrane was incubated with the following primary antibodies: mouse anti-TH (1:1000; Millipore) and mouse anti-GAPDH (1:5000; Abmart, Shanghai, China). After being washed with Tris-buffered saline containing 0.05% Tween-20, the membrane was incubated with an anti-mouse peroxidase-conjugated secondary antibody (1:3000; CWBIO). The membrane was then washed with Tris-buffered saline containing 0.05% Tween-20, and the Super Signal West Pico chemiluminescent substrate (Thermo Scientific) was used for detection.
3
Protein Extraction and Analysis in Zebrafish
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Zebrafish larvae at 5 dpf or adult tail fins were lysed in RIPA (CWBIO, China) with PMSF (Sigma-Aldrich). The proteins were quantified, and subjected to SDS-PAGE. They were then transferred onto a PVDF membrane. The primary antibodies are listed as follows: mouse anti- GAPDH (1:5000; Abmart, China), mouse anti- actin (1:1000; Millipore), rabbit anti- zHCRT (1:1000; produced by HuaBio, Hangzhou, China), and rabbit anti- zP2RY11(1:1000 produced by HuaBio, Hangzhou, China).
4
Protein Expression in Colorectal Cancer Cells
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Total protein was extracted from three infected CRC cell lines (HCT116, RKO, and MC38) using RIPA lysis buffer supplemented with 1% PMSF. Total protein was separated using a 10% sodium dodecyl sulfate‒polyacrylamide gel and then transferred to a PVDF membrane. The membranes were blocked with 5% skim milk for 2 h and then incubated with rabbit anti-C6orf15 (Proteintech, USA; 1:1000 dilution), rabbit anti-β-catenin (Proteintech, USA; 1:1000 dilution), rabbit anti-ZEB1 (Abmart, Shanghai; 1:1000 dilution), rabbit anti-E-cadherin (Abmart, Shanghai; 1:1000 dilution), rabbit anti-N-cadherin (Abmart, Shanghai; 1:1000 dilution), rabbit anti-Vimentin (Proteintech, USA; 1:1000 dilution), rabbit anti-ZO-1 (Abmart, Shanghai; 1:1000 dilution), mouse anti-GAPDH (Abmart, Shanghai; 1:3000 dilution), rabbit anti-CPT1A (Proteintech, USA; 1:1000 dilution), and rabbit anti-LaminB1 (Proteintech, USA; 1:1000 dilution). The membranes were then incubated with secondary antibodies for 2 h. We assessed the protein blotting results using an enhanced chemiluminescence (ECL) assay kit (Thermo Fisher Scientific, Rockford, IL, USA), and each experiment was performed three times.
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