Rabbit polyclonal antibody against MAP2 was provided by Dr. Itzhak Fischer, Drexel University College of Medicine, Philadelphia, PA (Nunez and Fischer, 1997 (link)). Other antibodies were purchased as follows: β3-tubulin rabbit polyclonal and mouse monoclonal antibody (Covance), β-tubulin cy3 conjugated monoclonal mouse antibody (Sigma), TPX2 rabbit polyclonal antibody (Santa Cruz Biotechnology), mouse monoclonal actin antibody (Abcam), mouse monoclonal GAPDH antibody (Abcam). Appropriate fluorescent goat secondary antibodies were purchased from Jackson Immunoresearch.
Gapdh mouse monoclonal antibody
Manufactured by Abcam
Sourced in United Kingdom
The GAPDH mouse monoclonal antibody is a primary antibody that specifically binds to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a widely expressed enzyme involved in glycolysis and other cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
β3 tubulin rabbit polyclonal and mouse monoclonal antibody, by Fortrea (2 mentions)
Appropriate fluorescent secondary antibodies, by Jackson ImmunoResearch (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Cd133, by Abcam (1 mentions)
Py705 stat3, by Abcam (1 mentions)
Ecl system, by Merck Group (1 mentions)
Stat3, by Abcam (1 mentions)
Atg7 rabbit polyclonal antibody, by Thermo Fisher Scientific (1 mentions)
Prestin goat polyclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Gp62 c, by Progen Biotechnik (1 mentions)
Ripa lysis buffer, by Fudebio (1 mentions)
Immobilon p pvdf membrane, by Merck Group (1 mentions)
Bolt 4 12 bis tris plus, by Thermo Fisher Scientific (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Pngase f, by New England Biolabs (1 mentions)
6 protocols using gapdh mouse monoclonal antibody
1
Immunofluorescence Staining of Neurons
2
Antibody Characterization for Neuronal Cytoskeleton
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Rabbit polyclonal antibody against MAP2 was provided by Itzhak Fischer (Drexel University College of Medicine, Philadelphia, PA; Nunez and Fischer, 1997 (link)). Other antibodies were purchased as noted: detyrosinated tubulin rabbit polyclonal antibody (Millipore), acetylated tubulin mouse monoclonal antibody (Sigma-Aldrich), tyrosinated tubulin YL1/2 rat monoclonal antibody (Accurate Scientific, Westbury, NY), β3-tubulin rabbit polyclonal and mouse monoclonal antibody (Covance, Princeton, NJ), β-tubulin fluorescein isothiocyanate–conjugated monoclonal mouse antibody (Sigma-Aldrich), kinesin-5 phosphorylated at Thr-926 rabbit polyclonal antibody (Phospho Solutions, Aurora, CO), total kinesin-5 rabbit polyclonal antibody (Abcam, Cambridge, UK), and mouse monoclonal GAPDH antibody (Abcam). Appropriate fluorescent secondary antibodies were purchased from Jackson Immunoresearch (West Grove, PA).
3
Western Blot Analysis of Protein Markers
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Total/cytoplasm/nucleus protein was extracted from tumor tissues, non-tumor adjacent tissues, or liver cancer cell lines using the Total/Cytoplasm/Nucleus Protein Extraction Kit (Solarbio, China). An equivalent of 50 µg of the protein extract was resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and electrotransferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). The membrane was blocked for 2 h at room temperature using milk (5%) was used to block membranes. Subsequently, the membranes were probed with primary antibodies, including rabbit polyclonal antibodies to AQP3 (1:2000, Abcam, USA), CD133 (1:1500, Abcam), JAK1 (1:1000, Abcam), pY-JAK1 (1:2000, Abcam), JAK2 (1:1500, Abcam), pY-JAK2 (1:1500, Abcam), STAT3 (1:2000, Abcam), pY705-STAT3 (1:2000, Abcam), GAPDH mouse monoclonal antibody (1:2000, Abcam), Histone H3 (phospho S10) rabbit monoclonal antibody (1:500, Abcam) overnight at 4 °C, followed by incubation with secondary antibodies for 2 h at room temperature. The immunoreactive bands were identified using an ECL system (Millipore, USA). Every tissue was evaluated three times using Western blotting.
4
Western Blot Analysis of Autophagic Markers
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Cochleae and other organs were dissected and transferred to the homogenizer tubes and mixed with RIPA lysis buffer (Fudebio, FD008) with a protease inhibitor cocktail (Roche, 11697498001). The supernatant from centrifuged homogenates was mixed with sodium dodecyl sulfate buffer (Beyotime, P0015L), boiled, electrophoresed, and blotted onto a 0.2-μm polyvinylidene difluoride membrane (Millipore, Immobilon ISEQ00010). The primary antibodies were: Atg7 rabbit polyclonal antibody (Thermo Fisher, PA5-35203, 1:500), Prestin goat polyclonal antibody (Santa Cruz, sc-22694, 1:400), GAPDH mouse monoclonal antibody (Abcam, ab9484, 1:1000), β-Actin rabbit IgG antibody (Abmart, P30002, 1:1000), LC3 rabbit polyclonal antibody (CST, 4108, 1:1000), and P62 Guinea Pig polyclonal antibody C-terminal specific (Progen, GP62-C, 1:500). The ECL Kit and horseradish peroxidase-conjugated antibodies were used for detection. Images were obtained by GE ImageQuant LAS4000.
5
Protein Extraction and Western Blot
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Total protein extracts were isolated with NP-40 buffer (1% (v/v) NP-40, 0.15 M NaCl, 10 mM EDTA, 10 mM NaN3, 10 mM Tris-HCl pH 8) containing cOmplete™ Protease Inhibitor Cocktail (Roche, Basel, Switzerland, Cat#116974980001). The protein extracts (20 μg) were separated on SDS PAGE (Bolt™ 4–12% Bis-Tris Plus; Invitrogen; Cat#NW04120BOX) and transferred to Immobilon™-P PVDF membrane (Merck Millipore; Darmstadt, Germany, Cat#IPVH00010). Following the blocking in 5% (w/v) bovine serum albumin, the membranes were probed with primary antibodies FLAG-HRP (1:1000; Merck; Cat#F1804) or GAPDH mouse monoclonal antibody (1:5000; Abcam; Cambridge, UK, Cat#ab8245) and secondary antibody (1:5000; Merck Millipore; Cat#AP192P). To remove N-linked glycosylation moieties from cell lysates, equal quantities of protein were incubated with PNGase F (New England Biolabs; Ipswich, MA, USA, Cat#P0704S) at 37 °C for 24 h, prior to running on SDS-PAGE gels.
6
Protein Analysis of Rat Myocardial Tissue
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Myocardial tissue (0.5 g) from rats in each group was weighed and placed in a 1.5-mL EP tube. Tissue lysate (300 µL) was added. Then, the myocardial tissue was homogenized using the electric homogenate instrument. The sample was placed on ice for 20 min and centrifuged at 15,000 g for 10 min. The supernatant was used to detect the total protein concentration using the BCA protein ELISA kit from Biyuntian Co., Shanghai, China. Loading buffer (5X) was added according to the sample volume and boiled in water for 10 min. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was conducted according to the amount of the total protein. When bromophenol blue ran to the edge, the electrophoresis was stopped and the protein on SDS-PAGE was transferred to a polyvinyl difluoride (PVDF) membrane. The PVDF membrane was incubated with 5% skim milk powder for 30 min, coated overnight with RIP1 and RIP3 rabbit polyclonal antibody (Abcam, Cambridge, UK) and GAPDH mouse monoclonal antibody (Abcam). The membrane was washed with PBS three times the next day (5 min/wash), labeled with HRP sheep anti rabbit secondary antibody (ZSGB-BIO, Beijing, China) at room temperature for 1 h, and washed with PBS 3 times (5 min/wash). The sample was photographed and scanned after the illuminating agent was added.
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