Hrp anti rabbit igg

Manufactured by CWBIO

Sourced in United States

HRP-anti-rabbit IgG is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and visualize rabbit primary antibodies in various immunoassay techniques.

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2 protocols using hrp anti rabbit igg

Immunoblotting of Cell Signaling Proteins

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The procedures were described as previous[20]. Briefly, equal amounts of cell protein per sample were electrophoresed through SDS-PAGE (12% polyacrylamide), and then were transferred onto a PVDF membrane (Roche, USA). Blots were incubated overnight with primary antibodies listed below. The primary antibodies included ERK1/2, p-ERK1/2, JNK, p-JNK, PI3K, p-PI3K (Tyr458), AKT, p-AKT, P38, p-P38, P65, p-P65, IκBα, p-IκBα (Cell Signaling Technology, USA), and GAPDH (CWBIO, China). Then, after incubation with secondary HRP-anti-rabbit IgG or HRP-anti-mouse IgG (CWBIO), the PVDF membrane were visualized with the ECL luminescence reagent (Thermo Fisher Scientific, USA). The immunoreactive bands were quantified using ImageJ.

Chen Y., Fu W.L., Gan X.D., Xing W.W., Xia W.R., Zou M.J., Liu Q., Wang Y.Y., Zhang C, & Xu D.G. (2018). SAK-HV Promotes RAW264.7 cells Migration Mediated by MCP-1 via JNK and NF-κB Pathways. International Journal of Biological Sciences, 14(14), 1993-2002.

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Protein Extraction and Western Blot Analysis

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Ventricles from 4-HT-treated Tg(cmlc2:creER;cmlc2:nRSGG) and Tg(cmlc2:nRSGG) animals, as well as grl^5nt^−/− and WT sibling zebrafish at 7 dpa were collected and homogenized with an electric homogenizer in cold RIPA lysis buffer (50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton X-100 and 0.1% SDS) supplemented with proteinase inhibitors (Millipore) and phosphatase inhibitor cocktail (Apex Bio). The lysates were loaded into 12.5% SDS-PAGE after boiling for 5 min in SDS-PAGE sample loading buffer. Proteins were then transferred to PVDF membranes using a Trans-Blot Turbo Transfer System (Bio-Rad). Signals were detected by the Clarity Western ECL Substrate (Bio-Rad) and scanned using a ChemiScope series system (Clinx). Antibodies used for western blot in this study were as follows: anti-Smyd2 (Cell Signaling; #9734; 1:500), anti-P-Stat3 (Santa Cruz; sc-8059; 1:500), anti-Stat3 (Santa Cruz; sc-8019; 1:500), anti-methyl-lysine (Abbkine; ABM0060; 1:1000), anti-GAPDH (Abcam; ab181602; 1:10,000), anti-GAPDH (Abmart; M20006L; 1:4000), anti-actin (Sigma; A5441; 1:4000), anti-Flag (Sigma; 7425/1804; 1:4000), anti-Myc (HuaAn Biotechnology; R1208-1; 1:4000), anti-HA (Santa Cruz; sc-805; 1:4000), HRP-anti-rabbit IgG (Cwbio; CW0103S; 1:5000) and HRP-anti-mouse IgG (Cwbio; CW0102S; 1:5000).

She P., Zhang H., Peng X., Sun J., Gao B., Zhou Y., Zhu X., Hu X., Lai K.S., Wong J., Zhou B., Wang L, & Zhong T.P. (2020). The Gridlock transcriptional repressor impedes vertebrate heart regeneration by restricting expression of lysine methyltransferase. Development (Cambridge, England), 147(18), dev190678.

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