Alexa fluor 680 conjugated anti rabbit igg

Manufactured by Thermo Fisher Scientific

Alexa Fluor 680-conjugated anti-rabbit IgG is a secondary antibody that binds to rabbit primary antibodies. It is conjugated to the fluorescent dye Alexa Fluor 680, which can be detected using appropriate instrumentation.

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Lab products found in correlation

Bcip nbt, by Beyotime (1 mentions) Anti phospho p38 mapk antibody, by Cell Signaling Technology (1 mentions) Anti p38 mapk, by Cell Signaling Technology (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Lysis buffer, by Beyotime (1 mentions) Irdye 800 conjugated anti mouse igg, by Rockland Immunochemicals (1 mentions) Mouse β actin, by Santa Cruz Biotechnology (1 mentions)

2 protocols using alexa fluor 680 conjugated anti rabbit igg

Quantification of p38 MAPK Activation

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Intestinal tissue was isolated and homogenized in lysis buffer (Beyotime Biotechnology, China) on ice. The homogenate was centrifuged (14 000 rpm, 15 min, 4°C) and the level of total supernatant protein was measured by BCA protein assay kit (Beyotime Biotechnology, China). The protein samples (20 μg) were boiled in Laemmle’s sampling buffer and electrophoresed on 10% sodium dodecyl sulfate-polyacrylamide gels. Next, the proteins were transferred to a polyvinylidene difluoride membrane and blocked with Tris-buffered saline containing 5% bovine serum albumin and 0.1% Tween 20. They were then incubated with anti-p38 MAPK or the anti-phospho-p38 MAPK antibody (1: 1000; Cell Signaling Technology, Danvers, MA) for 16 h at 4°C, followed by incubation with Alexa Fluor 680-conjugated anti-rabbit IgG (Molecular Probes). After washing 5 times for 5 min each time, the membranes were visualized by NBT/BCIP (Beyotime Institute of Biotechnology, Nantong, China). The protein bands were scanned and quantified using ImageJ software version 1.34 s (Wayne Rasband National Institute of Health) using β-actin for normalization.

Wan Y.D., Zhu R.X., Bian Z.Z, & Pan X.T. (2019). Improvement of Gut Microbiota by Inhibition of P38 Mitogen-Activated Protein Kinase (MAPK) Signaling Pathway in Rats with Severe Acute Pancreatitis. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 4609-4616.

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Cell culture protocol for glioblastoma, colon cancer, and normal fibroblasts

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Human glioblastoma cells Gli36, human colon carcinoma cells HCT116 and normal skin fibroblasts OSU2 were maintained according to established methods as described. Gli36 cell line was grown in DMEM supplemented with 10% fetal bovine serum (FBS) and antibiotics (50 units/ml penicillin and 50 μg/ml streptomycin). HCT116 cells were cultured in McCoy’s 5A medium supplemented with 10% FBS and antibiotics as above. Normal human skin OSU-2 fibroblasts were established and maintained in culture as described in previous studies (Venkatachalam et al., 1995 (link)). All cells were grown at 37°C in a humidified 5% CO₂ atmosphere. The DC protein quantitation reagents were from Bio-Rad. Antibodies against the following proteins were diluted in blocking buffer: mouse β-actin (Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1/3000; rabbit phospho-histone H2A.X, serine 139 (Millipore, Billerca, MA) diluted 1/3000. Alexa Fluor 680-conjugated anti-rabbit IgG (Molecular Probes) and IRDye800 conjugated anti-mouse IgG (Rockland, Gilbertsville, PA) were used as secondary antibodies. Texas red and FITC conjugated fluorescent antibodies were from Santa Cruz Biotechnology.

Rehmani N., Zafar A., Arif H., Hadi S.M, & Wani A.A. (2017). Copper-mediated DNA damage by the neurotransmitter dopamine and L-DOPA: A pro-oxidant mechanism. Toxicology in vitro : an international journal published in association with BIBRA, 40, 336-346.

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