Briefly, the NSCLC cell lines were lysed with RIPA buffer containing protease inhibitor. Following determination of the protein concentrations using the BCA Protein Assay kit (Beyotime Institute of Biotechnology, Haimen, China), equal amounts of protein per sample (20 µg) were separated on 10% sodium dodecyl sulfate (SDS) polyacrylamide gels for 2 h and transferred to nitrocellulose membranes. The membranes were subsequent blocked with 5% low-fat milk in TBST for 1 h and incubated with primary antibodies overnight at 4°C. The primary antibodies included: anti-HURP (1:2,000; Abcam) and GAPDH (1:2,000; Santa Cruz Biotechnology). This was followed by incubation with secondary antibodies (1:5,000; Santa Cruz Biotechnology) for 1.5 h at room temperature and rinsed four times with TBST for 8 min each. Reactions were visualized using the ECL system (Thermo Fisher Scientific, Waltham, MA, USA).
Anti hurp
Manufactured by Abcam
Anti-HURP is a laboratory reagent that recognizes and binds to the HURP (Hepatoma Upregulated Protein) protein. HURP is involved in the regulation of microtubule dynamics during cell division. Anti-HURP can be used to detect and study the HURP protein in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Ecl system, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ab18250, by Abcam (1 mentions)
Ab70744, by Abcam (1 mentions)
Anti mouse igg hrp, by Dianova (1 mentions)
Anti rabbit igg hrp, by Dianova (1 mentions)
Anti p y15 cdk1, by Cell Signaling Technology (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
F1804, by Merck Group (1 mentions)
Anti flag, by Merck Group (1 mentions)
2 protocols using anti hurp
1
Western Blot Analysis of HURP Protein
2
Kif18A Protein Expression and Antibody Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Full-length Xl_Kif18A was cloned from egg cDNA using primers 5′-attaggccggcccatggaagcctcccaagaggacgtc-3′ and 5′-taatggcgcgccctcagggatctttaagggctgc-3′. Rabbit polyclonal antibodies were generated against a C-terminal peptide: N-CGGRKKAALKDP-C (Ab18Apep), a protein fragment containing an N-terminal Serine: aa 845-953 (Ab18A-C) and a fragment from aa 1-104 tagged with an N-terminal His6-SMT3 (Ab18AN). Other antibodies used were: human Kif18AN (Mayr et al., 2007 (link); 1 µg/ml), anti-GFP (Covance, MMS-118P, 1:1000); anti-cyclin B2 (Abcam, ab18250; 1:1000), anti-XErp1 (Schmidt et al., 2005 (link); 1 µg/ml); anti- c-Mos (Santa Cruz, C237; 1:250), anti pY15-Cdk1 (Cell Signaling, 9111, 1:1000); anti-Flag (Sigma, F1804, 1:1000); anti α-tubulin (Sigma, F2168, 1:2000), anti HURP (Abcam, ab70744, 1:500), anti-mouse IgG-HRP and anti-rabbit IgG-HRP (Dianova, 115-035-062 and 111-035-003, both 1:3000).
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