Log In Sign up
The largest database of trusted experimental protocols

Monoclonal anti gapdh

Manufactured by Merck Group
Visit Supplier
Sourced in United States, United Kingdom

Monoclonal anti-GAPDH is a laboratory reagent used for the detection and quantification of the enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in biological samples. This antibody is specific to the GAPDH protein and can be used in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry.

Automatically generated - may contain errors

Lab products found in correlation

Las 4000 chemiluminescence detection system, by Fujifilm (2 mentions) Polyclonal anti 14 3 3ζ, by Santa Cruz Biotechnology (2 mentions) Horseradish peroxidase hrp linked anti rabbit, by Vector Laboratories (1 mentions) Monoclonal anti β actin, by Abcam (1 mentions) Alexa fluor 680, by Thermo Fisher Scientific (1 mentions) Ir800dye, by Rockland Immunochemicals (1 mentions) Protein assay, by Bio-Rad (1 mentions) Rabbit anti cox 4, by Cell Signaling Technology (1 mentions) Pvdf blotting membrane, by Merck Group (1 mentions) 6 well culture plates, by Avantor (1 mentions) Rabbit anti β tubulin, by Cell Signaling Technology (1 mentions) Rabbit anti grp78, by Merck Group (1 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Image studio software, by LI COR (1 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Ecl chemiluminescence system, by Thermo Fisher Scientific (1 mentions) Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Fusion solo s, by Vilber (1 mentions) Anti 14 3 3ζ, by Santa Cruz Biotechnology (1 mentions) Anti hnrnp q, by Merck Group (1 mentions)

7 protocols using monoclonal anti gapdh

1

Western Blot and Slot Blot Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein samples (20 μg) were used for western blot and slot blot analysis. The following primary antibodies were used: polyclonal anti-LRRK2 (1:2,500; Millipore), monoclonal anti-CD45 (1:2,500; Abcam, Cambridge, MA, USA), polyclonal anti-MMP-8 (1:2,500; Abcam), monoclonal anti-β-actin (1:2,500; Abcam), and monoclonal anti-GAPDH (1:2,500; Millipore). We used horseradish peroxidase (HRP)-linked anti-rabbit (1:2,500; Vector Laboratories, Burlingame, CA, USA) or HRP-linked anti-mouse (1:2,500; Vector Laboratories) IgG antibodies as secondary antibodies. After the incubation with primary and secondary antibodies, the blotted membrane was developed using ECL prime (Merck, Boston, MA, USA). Intensity of each band was quantified using Image J program (version 1.46r; NIH), and normalized to the expression level of β-actin or GAPDH as loading control.
Kim T., Jeon J., Park J.S., Park Y., Kim J., Noh H., Kim H.S, & Seo H. (2021). Matrix Metalloproteinase-8 Inhibitor Ameliorates Inflammatory Responses and Behavioral Deficits in LRRK2 G2019S Parkinson’s Disease Model Mice. Biomolecules & Therapeutics, 29(5), 483-491.
+ Open protocol
+ Expand
2

Immunoblotting of Cardiac Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse cardiac homogenates were prepared as described, subjected to electrophoresis on 6% (for RyR2) or 12% acrylamide gels (for CaMKII and PLN), and transferred onto polyvinyl difluoride (PVDF) membranes.[28 (link)] Membranes were probed with monoclonal anti-RyR2 (1:5,000; Thermo Fisher Scientific, Waltham, MA), polyclonal anti-pS2814-RyR2 (1:1,000),[17 (link)] polyclonal anti-pS2808-RyR2 (1:1,000),[28 (link)] monoclonal anti-pT286-CaMKII (1,1,000; Cayman Chemicals, Ann Arbor, MI), polyclonal anti-CaMKII (1:1,000; custom made for our lab), polyclonal anti-pT17-PLN (1:5,000; Badrilla, Leeds, UK), monoclonal anti-PLN (1:5,000; Badrilla, Leeds, UK), and monoclonal anti-GAPDH (1:10,000; Millipore, Temecula, CA) antibodies. Membranes were then incubated with secondary anti-mouse and anti-rabbit antibodies conjugated to Alexa-Fluor 680 (Invitrogen Molecular Probes, Carlsbad, CA) and IR800Dye (Rockland Immunochemicals, Gilbertsville, PA), respectively, and bands were quantified using infrared visualization (Odyssey System, Lincoln, NE) and densitometry (ImageJ Data Acquisition Software, National Institute of Health, Bethesda, MD). GAPDH was used as a loading control.
Respress J.L., Gershovich P.M., Wang T., Reynolds J.O., Skapura D.G., Sutton J.P., Miyake C.Y, & Wehrens X.H. (2014). Long-Term Simulated Microgravity Causes Cardiac RyR2 Phosphorylation and Arrhythmias in Mice. International journal of cardiology, 176(3), 994-1000.
+ Open protocol
+ Expand
3

Protein Extraction and Western Blot Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells grown to confluence on 6-well culture plates (VWR) were harvested after the specified treatment period and washed with PBS. Protein was extracted from the cells using mammalian protein extraction reagent with protease inhibitor cocktail (Pierce Biotechnology, Rockford, IL), and quantified with a protein assay (Bio-Rad, Hercules, CA). Equal amounts of protein were resolved on 8–16% Tris-HCL polyacrylamide gels (Pierce Biotechnology, Rockford, IL) and transferred to PVDF blotting membranes (Millipore, Billerica, MA). Membranes were probed with rabbit polyclonal anti-gamma glutamyl cysteine ligase, catalytic subunit (GCLC) (Abcam, Cambridge, MA), rabbit anti-cleaved caspase 3 (Cell Signaling Technology, MA), mouse anti-CHOP (Pierce Biotechnology, IL), rabbit anti-GRP78 (Sigma Aldrich, St. Louis, MO), rabbit anti-COX IV (Cell Signaling Technology, MA), rabbit polyclonal anti-GRX2 (GeneTex, Irvine, CA) and rabbit anti-β-Tubulin (Cell Signaling Technology, MA) overnight at 4°C. After incubation with corresponding secondary antibody tagged with horseradish peroxidase, signals were detected using an ECL chemiluminescence system (Pierce Biotechnology, IL). Membranes were then stripped and reprobed with monoclonal anti-GAPDH (Millipore, Billerica, MA). Protein band intensity was measured by Image Studio Software (Li-Cor, Lincoln, NE) [30 (link)].
Matsunaga D., Sreekumar P.G., Ishikawa K., Terasaki H., Barron E., Cohen P., Kannan R, & Hinton D.R. (2016). Humanin Protects RPE Cells from Endoplasmic Reticulum Stress-Induced Apoptosis by Upregulation of Mitochondrial Glutathione. PLoS ONE, 11(10), e0165150.
+ Open protocol
+ Expand
4

Immunoblot Analysis of Circadian Clock Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblot analyses were performed using polyclonal anti-PER1, monoclonal anti-hnRNP Q (Sigma-Aldrich), monoclonal anti-PTBP1 (Abcam), monoclonal anti-YBX1 (Abcam), polyclonal anti-hnRNP K (Abcam), monoclonal anti-hnRNP A1 (Abcam), monoclonal anti-FLAG (Sigma-Aldrich), polyclonal anti-14-3-3ζ (Santa Cruz Biotechnology) and monoclonal anti-GAPDH (Millipore) as primary antibodies. Horseradish peroxidase-conjugated species-specific secondary antibodies (goat, Santa Cruz Biotechnology; guinea pig, Santa Cruz Biotechnology; mouse, Thermo Fisher Scientific; rabbit, Jackson ImmunoResearch Laboratories) were visualized by using a SUPEX ECL solution kit (Neuronex) or a PierceTM ECL Western Blotting substrate (Thermo Fisher Scientific) under LAS-4000 chemiluminescence detection system (FUJIFILM) and a FUSION Solo S (Vilber). The acquired images were analyzed according to the manufacturer’s instructions.
Kim W., Shin J.C., Lee K.H, & Kim K.T. (2020). PTBP1 Positively Regulates the Translation of Circadian Clock Gene, Period1. International Journal of Molecular Sciences, 21(18), 6921.
+ Open protocol
+ Expand
5

Immunoblotting for Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunoblotting, cells were disrupted with complete protein solubilizing buffer containing 1% SDS and 2M urea in PBS, followed by sonication. Immunoblot analyses were performed with polyclonal anti-CRY1, monoclonal anti-GAPDH (Millipore), polyclonal anti-14-3-3ζ (Santa Cruz Biotechnology), and polyclonal anti-hnRNP Q (Sigma-Aldrich) antibodies. Horseradish peroxidase-conjugated mouse (Thermo Scientific) and rabbit (Jackson ImmunoResearch Laboratories) secondary antibodies were visualized with SUPEX ECL reagent (Neuronex) and a LAS-4000 system (FUJI FILM), according to the manufacturer’s instructions. Acquired images were further analyzed with the Image Gauge program (FUJIFILM).
Lim I., Jung Y., Kim D.Y, & Kim K.T. (2016). HnRNP Q Has a Suppressive Role in the Translation of Mouse Cryptochrome1. PLoS ONE, 11(7), e0159018.
+ Open protocol
+ Expand
6

Immunoblot Analysis of Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblot analyses were performed using polyclonal anti-CRY1, monoclonal anti-AUF1 (Millipore), polyclonal anti-14-3-3ζ (Santa Cruz Biotechnology), monoclonal anti-RPS3 (Cell Signaling), polyclonal anti-RPS14 (Santa Cruz Biotechnology), polyclonal anti-EIF3B (Santa Cruz Biotechnology) and monoclonal anti-GAPDH (Millipore) as primary antibodies. Horseradish peroxidase-conjugated species-specific secondary antibodies (goat, Santa Cruz Biotechnology; guinea pig, Santa Cruz Biotechnology; mouse, Thermo Scientific; rabbit, Jackson ImmunoResearch Laboratories) were visualized using a SUPEX ECL solution kit (Neuronex) and a LAS-4000 chemiluminescence detection system (FUJIFILM), and the acquired images were analysed using Image Gauge (FUJIFILM) according to the manufacturer’s instructions.
Lee K.H., Kim S.H., Kim H.J., Kim W., Lee H.R., Jung Y., Choi J.H., Hong K.Y., Jang S.K, & Kim K.T. (2014). AUF1 contributes to Cryptochrome1 mRNA degradation and rhythmic translation. Nucleic Acids Research, 42(6), 3590-3606.
+ Open protocol
+ Expand
7

Western Blot Analysis of NKX2.5 Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were homogenized in lysis buffer containing 135 mM NaCl, 1 mM MgCl2, 2.7 mM KCl, 20 mM Tris, pH 8.0, 1% Triton, 10% glycerol and protease and phosphatase inhibitors (0.5 mM Na3VO4, 10 mM NaF, 1 mM leupeptin, 1 mM pepstatin, 1 mM okadaic acid, and 0.2 mM phenylmethylsulfonyl fluoride), and then syringed five times. An aliquot was used to determine the concentration of protein by BCA protein assay kit (Pierce, Rockford, IL, USA), as recommended by the manufacturer. Samples were then subjected to SDS/PAGE electrophoresis, transferred to PVDF membranes, and probed with the following antibodies: 1:2000 polyclonal anti-NKX2.5, Sigma-Aldrich; 1:4000 monoclonal anti-GAPDH, Millipore; 1:2000 anti-rabbit IgG HRP-linked antibody and 1:4000 anti-mouse IgG HRP-linked antibodies from Cell Signaling. Detection of the proteins was performed using ECL (Thermo Scientific, Rockford, IL, USA).
Penha R.C., Buexm L.A., Rodrigues F.R., de Castro T.P., Santos M.C., Fortunato R.S., Carvalho D.P., Cardoso-Weide L.C, & Ferreira A.C. (2018). NKX2.5 is expressed in papillary thyroid carcinomas and regulates differentiation in thyroid cells. BMC Cancer, 18, 498.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!