Gapdh g8795

Manufactured by Merck Group

Sourced in United States

GAPDH (G8795) is a laboratory equipment product offered by the Merck Group. It is an enzyme involved in the glycolytic pathway, catalyzing the conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate. This product is used for research purposes, though its specific applications are not elaborated on in this factual description.

Automatically generated - may contain errors

Lab products found in correlation

Ecl plus reagent, by Merck Group (5 mentions) Quantity one 1 d analysis software, by Bio-Rad (5 mentions) Rb2 p130 r27020, by BD (4 mentions) P27kip1 3686, by Cell Signaling Technology (4 mentions) P16ink4a ab54210, by Abcam (4 mentions) Rb1 av33212, by Merck Group (3 mentions) P21cip1 sc 397, by Santa Cruz Biotechnology (3 mentions) Rala 4799, by Merck Group (2 mentions) Ralb 90879, by Merck Group (2 mentions) β actin a1978, by Merck Group (2 mentions) Anti mouse igg hrp 7076, by Merck Group (2 mentions) Anti rabbit igg hrp 7074, by Merck Group (2 mentions) Mab374, by Merck Group (1 mentions) Tfeb a303 673 a antibody, by Fortis Life Sciences (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Poly 1 c, by InvivoGen (1 mentions) Sc 9996, by Santa Cruz Biotechnology (1 mentions) Sc 11386, by Santa Cruz Biotechnology (1 mentions) Poly adp ribose polymerase parp sc 7150, by Santa Cruz Biotechnology (1 mentions) Flag f1804, by Merck Group (1 mentions)

Close spelling variants of this product

GAPDH (1812 citations) G8795 (86 citations) Anti-GAPDH (G8795) (7 citations) Anti-GAPDH antibody (G8795) (2 citations)

19 protocols using gapdh g8795

Western Blot Immunodetection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

See Sizemore et al. [24] for western blotting and immunodetection protocol. Antibodies were obtained from Cell Signaling Technologies (RalA: 4799, RalB: 90879, β-actin: A1978, anti-mouse-IgG HRP: 7076, anti-rabbit IgG HRP: 7074) and Millipore Sigma (GAPDH: G8795 or MAB374).

Thies K., Cole M.W., Schafer R., Spehar J.M., Richardson D.S., Steck S.A., Das M., Lian A.W., Ray A., Shakya R., Knoblaugh S.E., Timmers C., Ostrowski M.C., Chakravarti A., Sizemore G.M., & Sizemore S.T. (2021). The Small G-Protein RalA Promotes Progression and Metastasis of Triple Negative Breast Cancer.

+ Open protocol

+ Expand

Western Blotting and Immunodetection

Check if the same lab product or an alternative is used in the 5 most similar protocols

See Sizemore et al. [24 (link)] for western blotting and immunodetection protocol. Antibodies were obtained from the Cell Signaling Technologies (RalA: 4799, RalB: 90879, β-actin: A1978, anti-mouse-IgG HRP: 7076, anti-rabbit IgG HRP: 7074) and Millipore Sigma (GAPDH: G8795 or MAB374).

Thies K.A., Cole M.W., Schafer R.E., Spehar J.M., Richardson D.S., Steck S.A., Das M., Lian A.W., Ray A., Shakya R., Knoblaugh S.E., Timmers C.D., Ostrowski M.C., Chakravarti A., Sizemore G.M, & Sizemore S.T. (2021). The small G-protein RalA promotes progression and metastasis of triple-negative breast cancer. Breast Cancer Research : BCR, 23, 65.

+ Open protocol

+ Expand

Antibody Reagents for FLCN Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total FLCN antibody (11236-2-AP) was purchased from Proteintech (Rosemont, IL, USA). Flag (F7425), α-tubulin (T6074), and GAPDH (G8795) were obtained from MilliporeSigma. AMPKα1 (07-350) and AMPKα2 (07-363) antibodies were obtained from MilliporeSigma. Acetyl–coenzyme A (CoA) carboxylase (ACC; 3676), phospho-ACC1 (Ser79; 3661), AMPKα (2532), phospho-AMPKα (Thr172; 2535), AMPKβ1 (4178), lamin A/C (4777), p70 S6 kinase (9202), phospho-p70 S6 kinase (Thr389; 9206), Raptor (2280), phospho-Raptor (Ser792; 2083), S6 ribosomal protein (5G10), phospho-S6 ribosomal protein (Ser235/236; 2211), and TFE3 (14779) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). TFEB (#A303-673A) antibody was purchased from Bethyl Laboratories (Montgomery, TX, USA). Horseradish peroxidase–conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). AMPKγ1 (TA300519) antibody was from OriGene Technologies (Rockville, MD, USA). A site-specific rabbit polyclonal antidody against phospho-TFEB (Ser142) was generated by YenZym Antibodies (South San Francisco, CA, USA) by immunization and affinity purification with a phosphorylated peptide of the sequence identical between human and mouse (i.e., PN-*S-PMAMLHIGSNPC-amide, where the asterisk denotes the phosphorylated residue).

Collodet C., Foretz M., Deak M., Bultot L., Metairon S., Viollet B., Lefebvre G., Raymond F., Parisi A., Civiletto G., Gut P., Descombes P, & Sakamoto K. (2019). AMPK promotes induction of the tumor suppressor FLCN through activation of TFEB independently of mTOR. The FASEB Journal, 33(11), 12374-12391.

+ Open protocol

+ Expand

Antibody Generation and Reagent Validation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rabbit polyclonal antibody against EV-A71 3A was prepared as described (Tang et al., 2007 (link)). Mouse monoclonal antibodies against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (G8795) and Flag (F1804) were purchased from Sigma-Aldrich (St Louis, MO, USA). Rabbit monoclonal antibodies against phospho-eIF2α (#3597) and PKR (Ab32052, against amino acids 50–150 of human PKR) were obtained from Cell Signaling (Beverly, MA, USA) and Abcam (Cambridge, UK), respectively. Mouse monoclonal antibody against green fluorescent protein (GFP) (SC-9996) and rabbit polyclonal antibodies against eIF2α (SC-11386) and poly (ADP-ribose) polymerase (PARP) (SC-7150) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse monoclonal antibody against EV-A71 3C and rabbit polyclonal antibody against phospho-PKR were obtained from GeneTex (Irvine, CA, USA). Poly(I:C) and 2-aminopurine (2-AP) was purchased from Invivogen (San Diego, CA, USA). Staurosporine was purchased from Sigma-Aldrich.

Chang Y.H., Lau K.S., Kuo R.L, & Horng J.T. (2017). dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication. Frontiers in Cellular and Infection Microbiology, 7, 284.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Cycle Regulators

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in a buffer containing 0.1% Triton (Bio-Rad, Irvine, CA, USA) for 30 min in ice. Then, 20 μg of each lysate was electrophoresed in a polyacrylamide gel and electroblotted onto a nitrocellulose membrane. We used the following primary antibodies: RB1 (AV33212) and GAPDH (G8795) were from Sigma-Aldrich (MO, USA), RB2/P130 (R27020) was from BD Biosciences (San Jose, CA, USA), p27^KIP1 (3686) was from Cell Signaling, p107 (sc-318), p53 (sc-126), and p21^CIP1 (sc-397) were from Santa Cruz Biotechnology (Dallas, TX, USA), and p16^INK4A (ab54210) was from Abcam (Cambridge, UK). Immunoreactive signals were detected with a horseradish peroxidase-conjugated secondary antibody (ImmunoReagents, Raleigh, NC, USA) and reacted with ECL plus reagent (Merck Millipore). All of the antibodies were used according to the manufacturer’s instructions. The mean value was quantified densitometrically using Quantity One 1-D analysis software (Bio-Rad).

Alessio N., Acar M.B., Demirsoy I.H., Squillaro T., Siniscalco D., Bernardo G.D., Peluso G., Özcan S, & Galderisi U. (2020). Obesity is associated with senescence of mesenchymal stromal cells derived from bone marrow, subcutaneous and visceral fat of young mice. Aging (Albany NY), 12(13), 12609-12621.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

SDS-PAGE and western blotting were performed as previously described. [28 (link)] Briefly, cell lysates were generated using RIPA buffer containing protease phosphatase inhibitor cocktail (Pierce). Cell lysates were subject to centrifugation and the protein content in supernatants was determined using the BCA Protein Assay Kit (Pierce). Proteins were resolved by 7.5%-12% SDS-PAGE, transferred to PVDF membrane and blocked with 5% bovine serum albumin. Proteins were detected with the following antibodies; cleaved Caspase 3 (MAB835) and hGRP-78 (MAB4846) from R&D Systems, BAX (ab32503) and Bcl-2 (ab7973) from Abcam, GAPDH (G8795) from Sigma Aldrich, p-PERK (sc-32577) from Santa Cruz Biotechnology, and Caspase-3 (9662), IRE-1 (3294S), CHOP (2895S), p-eIF2α (3597s), eIF2 (9722), PERK (3192S), p-P38 (9211), P38 (9212), p-Erk1/2 (9101), Erk (9102), p-SAPK-JNK (9251), and SAPK/JNK (9252) from Cell Signaling. ATF6 (MA5-16172) was purchased from Thermo Fisher Scientific. Blots were incubated with HRP-conjugated secondary antibodies followed by enhanced chemiluminescence (ECL) detection. Results were quantified by densitometry of digitized images using ImageJ software (NIH, Bethesda, MD, ver.1.43) and expressed as a ratio to a GAPDH loading control.

Banerjee A., Ahmed H., Yang P., Czinn S.J, & Blanchard T.G. (2016). Endoplasmic reticulum stress and IRE-1 signaling cause apoptosis in colon cancer cells in response to andrographolide treatment. Oncotarget, 7(27), 41432-41444.

+ Open protocol

+ Expand

Western Blot Analysis of Adipocyte Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in a buffer containing 0.1% Triton (Bio-Rad, Hercules, CA, USA) for 30 min in ice; 20 μg of each lysate was electrophoresed in a polyacrylamide gel and electroblotted into a nitrocellulose membrane using Biorad transblot apparatus. We used the following primary antibodies: UCP1 (sc518024, Santa Cruz, USA), LPL (MABS1270), and GAPDH (G8795) from Sigma-Aldrich (St. Louis, MO, USA), PPARG (PA3-821A) from Thermo Fischer Scientific (Waltham, Massachusetts, USA). The primary antibodies were added to a nitrocellulose membrane in a buffer solution (TTBS 0.01X and 3% free fat milk) and incubated overnight at 4 °C. Then, the membranes were washed three times in T-TBS 1X and incubated with a horseradish peroxidase-conjugated secondary antibody (ImmunoReagents, Raleigh, NC, USA) and reacted with ECL plus reagent (Merck Millipore, Burlington, MA, USA). All antibodies were used according to the manufacturer’s instructions. The mean value was quantified densitometrically using Quantity One^® 1-D analysis software (Bio-Rad, Hercules, CA, USA).

Di Maio G., Alessio N., Peluso G., Perrotta S., Monda M, & Di Bernardo G. (2022). Molecular and Physiological Effects of Browning Agents on White Adipocytes from Bone Marrow Mesenchymal Stromal Cells. International Journal of Molecular Sciences, 23(20), 12151.

+ Open protocol

+ Expand

Western Blot Antibody and Chemical Reagents

Check if the same lab product or an alternative is used in the 5 most similar protocols

The CDK4 (A304-225A) antibody was purchased from Bethyl Laboratories (Montgomery, TX). AXL (4566), CAMKII (4436), CDK6 (13331), MEK1/2 (8727), MET (8198), MERTK (4319), and SRC (2109) were obtained from Cell Signaling Technology (Danvers, MA). β-catenin (610153) and GSK3β (610201) antibodies were purchased from BD Biosciences (San Jose, CA). GAPDH (G8795) and β-tubulin (T7816) antibodies were obtained from Sigma-Aldrich (St. Louis, MO). Secondary antibodies were purchased from LI-COR Biosciences (Lincoln, NE): IRDye® 800CW Goat anti-Mouse IgG (925-32210), IRDye® 680LT Goat anti-Mouse IgG (925-68020), IRDye®800CW Goat anti-Rabbit IgG (925-32211), and IRDye® 680LT Goat anti-Rabbit IgG (925-68021). The following chemicals were purchased from Cayman Chemicals (Ann Arbor, MI): abemaciclib (17740), CHIR-99021 (13122), dasatinib (11498), and ribociclib (17666). BMS-777607 (S1561) and palbociclib (S1579) were ordered from Selleck Chemicals (Houston, TX). Recombinant Wnt3A (315-20) was purchased from PeproTech (Rocky Hill, NJ).

Cousins E.M., Goldfarb D., Yan F., Roques J., Darr D., Johnson G.L, & Major M.B. (2017). Competitive Kinase Enrichment Proteomics Reveals that Abemaciclib Inhibits GSK3β and Activates WNT Signaling. Molecular cancer research : MCR, 16(2), 333-344.

+ Open protocol

+ Expand

Western Blot Analysis of Angiogenic Factors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were electrophoresed using a NuPAGE 10% Bis‐Tris gel (Invitrogen) and transferred to polyvinylidene membranes (Bio‐Rad Laboratories). Immunoblot detection was performed for vascular endothelial growth factor (VEGF) (ab46154, 1:1000; Abcam, Cambridge, MA), VEGFR2 (55B11, 1:1000; Cell Signaling, Danvers, MA), eNOS (1:1000; BD Bioscience, BD Bioscience, San Jose, CA) in mouse tissue, GSNOR in (1:1000, 11051‐1‐AP, Proteintech, Rosemont, IL) in human tissues, B‐actin (4957S, 1:1000, Cell Signaling, Danvers, MA), GAPDH (G8795, 1:1000, Sigma, St Louis, MO) and subsequent reaction with a goat anti‐rabbit horseradish peroxidase–conjugated antibody (1:1000; Cell Signaling). Then, membranes were developed by enhanced chemiluminescence (Super Signal West Pico, Thermo Scientific, Hampton, NH) and analyzed by the QuantityOne software (Bio‐Rad, Hercules, CA).

Kulandavelu S., Dulce R.A., Murray C.I., Bellio M.A., Fritsch J., Kanashiro‐Takeuchi R., Arora H., Paulino E., Soetkamp D., Balkan W., Van Eyk J.E, & Hare J.M. (2022). S‐Nitrosoglutathione Reductase Deficiency Causes Aberrant Placental S‐Nitrosylation and Preeclampsia. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 11(5), e024008.

+ Open protocol

+ Expand

Western Blot Protocol for Protein Detection

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Unless otherwise stated, all Western blotting was performed using equipment from BioRad as previously described [30 (link)]. Primary antibodies were used to detect BCRP (BXP-53, 1:5000; Enzo Life Sciences, Farmingdale, NY), β-actin (ab8227, 1:2000, Abcam, Cambridge, MA), transferrin receptor 1 (TFR-1, ab108985, 1:5000 Abcam), placental alkaline phosphatase (PLAP, ab133602, 1:10,000, Abcam), multidrug resistance-associated protein 1 (MRP1, ab3368, 1:2000, Abcam), cluster of differentiation 34 (CD34, ab81289 1:10,000, Abcam), histone H2A (25785, 1:1000, Cell Signaling, Danvers, MA), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, G8795, 1:1000, Sigma-Aldrich). HRP-linked secondary antibodies (anti-rabbit, anti-rat or anti-mouse, 1:2000 Sigma-Aldrich) were used to detect primary antibodies. After the addition of a Luminata Forte Western HRP substrate (Millipore, Billerica, MA), chemiluminescent protein-antibody complexes were visualized using a Fluorchem Imager (ProteinSimple, Santa Clara, CA). Semi-quantitative analysis of bands of the blots was performed using AlphaView Software (ProteinSimple).

Szilagyi J.T., Vetrano A.M., Laskin J.D, & Aleksunes L.M. (2017). Localization of the Placental BCRP/ABCG2 Transporter to Lipid Rafts: Role for Cholesterol in Mediating Efflux Activity. Placenta, 55, 29-36.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!