Total proteins from H9c2 cells and working heart tissues were extracted using RIPA lysis buffer, separated by electrophoresis on an SDS-polyacrylamide gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Billerica, MS, USA). The membrane was blocked with 5% nonfat milk in Tris-buffered saline/Tween 20 (TBST) and was incubated overnight with the following primary antibodies: anti-ox-CaMKIIδ (Met281/282) (1 : 1000, GeneTex), anti-p-CaMKIIδ (Thr286) (1 : 1000, Abcam), anti-CaMKIIα (1 : 10000, Abcam), anti-CaMKIIβ (1 : 500, Abcam), anti-CaMKIIδ (1 : 1000, Abcam), anti-CaMKIIγ (1 : 500, Abcam), anti-p-JNK1/2 (T183/Y185) (1 : 1000, CST), anti-JNK1/2 (1 : 1000, CST), anti-p-NF-κB p65 (Ser536) (1 : 1000, Abcam), anti-NF-κB p65 (1 : 1000, Abcam), anti-Bcl-2 (1 : 500, Abcam), anti-Bax (1 : 500, Abcam), anti-cytochrome c (1 : 1000, Abcam), and anti-GAPDH (1 : 1000, ZSGB-Bio, Beijing, China). The membrane was then washed and probed with horseradish peroxidase-conjugated goat anti-rabbit or anti-mouse antibody (1 : 8000, ZSGB-Bio) to enhance the chemiluminescence that was detected using a Fusion-FX6 imaging system (Vilber Lourmat, Marne-la-Valle, France).
Anti gapdh
Manufactured by ZSGB-BIO
Sourced in China, United States
The Anti-GAPDH is a laboratory reagent used for the detection and quantification of the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein in biological samples. GAPDH is a widely used housekeeping gene and its expression is commonly assessed in various experimental settings. The Anti-GAPDH provides a reliable and specific tool for researchers to study GAPDH expression levels.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (4 mentions)
Anti n cadherin, by Cell Signaling Technology (3 mentions)
Anti β actin, by ZSGB-BIO (3 mentions)
Anti myc, by Cell Signaling Technology (3 mentions)
Anti runx2, by Cell Signaling Technology (3 mentions)
Anti cyclin d1, by Cell Signaling Technology (3 mentions)
Anti β actin, by Merck Group (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Opti mem reduced serum medium, by Thermo Fisher Scientific (2 mentions)
Bca assay kit, by Beyotime (2 mentions)
Goat anti mouse rabbit igg horseradish peroxidase conjugated antibody, by Bio-Rad (2 mentions)
Trypsin, by Beyotime (2 mentions)
Anti murf1, by GeneTex (2 mentions)
Gtx110475, by GeneTex (2 mentions)
Anti p70s6k, by Cell Signaling Technology (2 mentions)
Anti phospho p70s6k thr389, by Cell Signaling Technology (2 mentions)
L carnitine, by Merck Group (2 mentions)
Ab12162, by Abcam (2 mentions)
Ab227385, by Abcam (2 mentions)
Ab154768, by Abcam (2 mentions)
41 protocols using anti gapdh
1
Western Blot Analysis of Cardiac Proteins
2
Western Blot Analysis of Lung Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse lung tissues or cells were lysed with RIPA Lysis Extraction Buffer (Beyotime Technology) along with protease inhibitor cocktail (Selleck). The total protein concentration was determined by a BCA protein assay reagent kit (Applygen Technologies Inc.) according to the manufacturer’s protocol. Total protein (20 µg) was loaded and separated by 10% SDS‒PAGE and then transferred to a PVDF membrane. Membranes were blocked with 5% skim milk in TBST buffer for 1 h and then incubated with primary antibodies overnight at 4 °C, followed by incubation with horseradish peroxidase-conjugated secondary antibodies. The following antibodies were used: anti-ZNF451 (Proteintech), anti-α-SMA (BOSTER), anti-Col1 (Abcam), and anti-GAPDH (ZSGB BIO). The signaling was visualized using a ChemiDocTM XRS + with Image LabTM Software (Bio-Rad, Hercules, California, USA) with an ECL kit (Tanon).
3
Molecular Signaling Regulation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
L-carnitine was obtained from Sigma Aldrich (St. Louis, MO, USA). The anti-AKT (1:1000, ab227385), antiphospho-AKT1 (Ser473) (1:1000, ab81283), anti-FOXO3a (1:1000, ab12162), and anti-phospho-FOXO3a (Ser253) (1:1000, ab154768) antibodies were purchased from Abcam Corp. (Cambridge, MA, USA). Anti-p70S6K (1:1000,2217s) and anti-phospho-p70S6K (Thr389)(1:1000, 4858s) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-MAFbx (1:1000, EB09088) was purchased from Everest Biotech. (Shanghai, China). Anti-MuRF1 (1:1000, GTX110475) was purchased from GeneTex (Irvine, CA, USA). The anti-GAPDH (1:1000, TA309157), anti-mouse and anti-rabbit secondary antibodies (1:5000-1:1000) were purchased from ZSGB-BIO (Beijing, China). Goat anti-mouse/rabbit IgG horseradish peroxidase-conjugated antibodies were purchased from Bio-Rad (Hercules, CA, USA).The trypsin and BCA assay kit were purchased from Beyotime (Shanghai, China). Lipofectamine2000 was purchased from Invitrogen (Carlsbad, CA, USA) and OPTI-MEM Reduced Serum Medium were purchased from Gibco (Langley, OK).The ELISA kit was purchased from Abebio (Wuhan, China).
4
Selective SHP-1 Inhibition Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NSC-87877 (565851), a SHP-1 selective inhibitor, was purchased from Calbiochem (Millipore, Billerica, MA, USA). The following antibodies were purchased: anti-His antibody from MBL (Nagoya, Japan), anti-Myc antibody from Applygen (Beijing, China), anti-PDZK1 antibody and anti-phosphor-SHP-1 (Tyr536) antibody from Abcam (Cambridge, UK), anti-PLCβ3 and anti-SHP-1 from Santa Cruz (Dallas, TX, USA), anti-FLAG M2 antibody and FLAG M2-agarose from Sigma (St Louis, MO, USA), anti-STAT5, anti-phosphor-STAT5 (Tyr694), anti-STAT3, anti-phosphor-STAT3 (Tyr705), anti-phosphor-Akt (Ser473) and anti-Akt from CST (Danvers, MA, USA), HRP-conjugated secondary antibodies and anti-GAPDH from ZSGB-BIO (Beijing, China).
5
Cholesterol Homeostasis and Wnt Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW1116, SW480 were originally obtained from ATCC and reserved in our laboratory. Filipin III (Santa Cruz), Bodipy 493/503 (Thermo Fisher), Cholesterol, β-Cyclodextrin, and AOM (Sigma), Avasimibe and LGK974 (Selleck), Nystatin (MCE), DSS (MP Biomedicals). Antibody against YAP, pYAPSer127, β-Catenin, c-Myc, FZD8, RhoA, c-Jun, GST were all from Abcam. Antibody against LRP6, pLRP6Ser1490, GSK-3β, pGSK-3βSer9, pLATS1Ser909, LATS1 were from CST. Antibody against pc-JunSer63, FZD1, CYR61 were from Sangon Biotech. Antibody against LDLR, HMGCR, SOAT2, FZD2, FZD5, FZD7, Myc were from Proteintech. Anti-SOAT1 was from Abclonal. Anti-GAPDH was from ZSGB-Bio.
6
Antibody characterization for research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used the following antibodies: anti-PTBP1 (1:1000 for Western blotting, 101043-T46; Sino Biology, China), anti-N-cadherin (1:1000 for Western blotting and 1:200 for immunofluorescence analysis, 13116 s; Cell Signaling Technology, Danvers, MA, USA), anti-TGF-β (1:1000 for Western blotting and 1:200 for immunofluorescence analysis, #3711; Cell Signaling Technology), and anti-GAPDH (1:1000 for Western blotting and 1:200 for immunofluorescence analysis, TA-08; ZSGB-BIO, Beijing, China).
7
Protein Expression Analysis of HXR9 and CXR9 Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with 60 μmol/L HXR9 or CXR9 for 2 hours. Total proteins were extracted by using RIPA lysis buffer with protease inhibitor cocktail and separated by SDS‐PAGE, blotted onto PVDF, then immunoreacted with primary antibody overnight at 4°C. The primary antibodies used were anti‐PBX (sc‐28313 at 1:200; Santa Cruz Biotechnology, Dallas, TX, USA), anti‐HOXB7 (ab51237 at 1:50; Abcam, Cambridge, MA, USA), anti‐HOXC6 (ab151575 at 1:1000; Abcam), anti‐HOXC8 (ab86236 at 1:1000; Abcam), anti‐Caspase‐3 (#9662 at 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti‐poly ADP ribose polymerase (anti‐PARP, #5625 at 1:1000; Cell Signaling Technology), anti‐c‐FOS (sc‐447 at 1:200; Santa Cruz Biotechnology), anti‐PI3K (#4249 at 1:1000; Cell Signaling Technology), anti‐AKT (#9272 at 1:1000; Cell Signaling Technology), anti‐p‐AKT (#5012 at 1:1000; Cell Signaling Technology), anti‐signal transducer and activator of transcription‐6 (anti‐STAT6, #5397 at 1:1000; Cell Signaling Technology), anti‐p‐STAT6 (#9364 at 1:1000; Cell Signaling Technology) and anti‐GAPDH (ZS‐25778 at 1:2000; ZSGB‐BIO, Beijing, China). Goat antirabbit IgG (ZB‐2301 at 1:5000; ZSGB‐BIO) and goat antimouse IgG (ZB‐2305 at 1:5000; ZSGB‐BIO) were used as the secondary antibodies.
8
Antibody Sourcing and Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The anti-His antibody was purchased from MBL (Nagoya, Japan). Anti-MAGI3, anti-Cyclin D1 and anti-GFAP antibodies from Abcam (Cambridge, UK); Anti-β-catenin and anti-GFP antibodies from Cell Signaling Technology (Beverly, CA); Anti-Flag M2 antibody and anti-Flag M2 affinity gels from Sigma (St Louis, MO); Anti-β-actin, anti-GAPDH and HRP-conjugated secondary antibodies from ZSGB-BIO (Beijing, China).
9
Protein Expression Analysis of Treated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After different treatment the cells were collected and washed in ice cold PBS, added with Cocktail cell lysis solution (Roche Molecular Bio chemicals, Indianapolis, IN, USA) and 1% phosphatase inhibitors (Kaiji Biological Technology, Nanjing, China) and lysed on ice for 40 minutes. The cell lysates were then centrifuged at 12000 RPM for 20 min at 4°C and the supernatant collected. The protein levels of all samples were quantified by a BCA assay according to the manufacturer's instructions (Kangwei Century Biotechnology, Beijing, China). Protein samples were equalized and separated by SDS/PAGE and transferred to PVDF membrane (Millipore) and detected by chemiluminescent HRP (horseradish peroxidase) substrate (Millipore). The antibodies used were as following: anti-Cleaved-PRAP1 (Cell Signaling Technology); anti-[acetyl-histone H3 (Lys9)] (Abcam); anti-GAPDH (ZSGB Bio, Beijing, China); anti-[acetyl-histone H3] (Abcam).
10
Reversing Immortalization in iDP6 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary DP cells and iDP6 cells were cultured. The iDP6 cells were treated with AdGFP (adenovirus with the ability to express GFP protein), AdFlip (adenovirus with the ability to express flip recombinase, which can interact with FRT thus remove the expression of SV40) or PBS. Forty-eight hours later, cells were collected and total proteins were extracted with RIPA lysis buffer (Beyotime, Shanghai, China). Then, total proteins were loaded to 1% SDS-PAGE gel (Beyotime, China) and transmitted to PVDF membrane (Bio-Rad, Hercules, CA, USA). The PVDF membrane were incubated with anti-SV40 (1:1,000; Santa Cruz Biotechnology, Dallas, TX, USA) and anti-GAPDH (1:500; ZSGB-bio, Beijing, China) antibodies. HRP labelled secondary antibodies were used, and the results were observed under ChemiDoc™ Touch Imaging System (Bio-Rad, Hercules, CA, USA). The experiment on reversing immortalization was performed twice.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!