Anti his tag

Manufactured by Abmart

Sourced in China

The Anti-His-tag is a laboratory equipment used for the detection and purification of proteins with a histidine (His) tag. It functions by specifically binding to the His-tag, which is a commonly used affinity tag for recombinant protein expression and purification.

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Lab products found in correlation

Anti flag tag, by GenScript (1 mentions) Anti ha tag, by Abmart (1 mentions) Goat anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions) Hoechst 33342, by Merck Group (1 mentions) Anti flag magnetic beads, by Selleck Chemicals (1 mentions) Ecl western blotting substrate, by Vazyme (1 mentions) Anti tsg101, by Proteintech (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Anti lpcat1, by Proteintech (1 mentions) Anti gm130, by Proteintech (1 mentions) Ecl reagent, by Solarbio (1 mentions) Hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions) Anti egfr, by Proteintech (1 mentions) Anti cd63, by Proteintech (1 mentions) Anti β actin, by Merck Group (1 mentions) Ripa lysis buffer, by Solarbio (1 mentions) Bca protein assay kit, by Solarbio (1 mentions)

Spelling variants of this product

Anti-His Tag antibody (5 citations) His-Tag (3 citations)

4 protocols using anti his tag

Immunofluorescence Staining Protocol

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Cells were washed in PBS and fixed in 4% paraformaldehyde for 30 min. After 3 washes in DPBS, cells were permeabilized with 0.3% Triton-X-100 for 60 min, blocked with PBS containing 2% BSA for 60 min and incubated with primary antibodies overnight at 4 °C. Then, the cells were treated with secondary antibody for 60 min at room temperature and mounted with Hoechst 33,342 (Sigma, Beijing, China) after 3 washes in DPBS.
The following antibodies were used in this study: anti-Myc-Tag (Abmart, Shanghai, China, A#M20002), anti-HA-Tag (Abmart, Shanghai, China, #M20003), anti-His-Tag (Abmart, Shanghai, China, #M20001), anti-Flag-Tag (Genscript, Nanjing, China, #A00170) and anti-dCas9 (Genscript, Nanjing, China, #A01885). Goat anti-rabbit secondary antibody (1:5000) (Santa Cruz, Dallas, TX, USA, #sc-2004). Samples were assessed by fluorescence microscopy at 530 nm and 480 nm excitation wavelengths (Olympus BX51, Olympus, Tokyo, Japan).

Jiang C., Geng L., Wang J., Liang Y., Guo X., Liu C., Zhao Y., Jin J., Liu Z, & Mu Y. (2023). Multiplexed Gene Engineering Based on dCas9 and gRNA-tRNA Array Encoded on Single Transcript. International Journal of Molecular Sciences, 24(10), 8535.

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Protein-Protein Interaction Assay

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All possible interactions between proteins in vitro were examined using pull-down assay^{45 (link)}. wspR was cloned into pET30a with a C-terminal His-tag, while rpfG was cloned into pET30a fused to a C-terminal FLAG-tag. Both WspR and RpfG were expressed in E. coli BL21 (DE3) and induced by 0.8 mM IPTG. 1 mL of bacteria lysate containing WspR-His or RpfG-FLAG was then incubated with 10 μL of anti-FLAG magnetic beads (Bimake, Shanghai, China). After overnight incubation at 4 °C, the beads were washed 3 times for 5 min each with 1 ml of 10 mM PBS buffer (pH, 7.5) containing 1% Triton X-100. Proteins bound to the beads were eluted with 45 μl elution buffer (0.2 M glycine HCl, pH 3.0), followed by the addition of 5 μl neutralization buffer (1 M Tris, pH 10). The eluted samples were boiled in 4×SDS loading buffer for 8 min. These samples were loaded into SDS-PAGE for Western blotting, and proteins were detected using anti-FLAG (No. M20008S, Abmart) and anti-His-tag (No. M30111L, Abmart) from Shanghai, China. Uncropped and unprocessed scans of gels & blots were provided in Supplementary Fig. 13.

Xu K., Wang L., Xiong D., Chen H., Tong X., Shao X., Li T, & Qian G. (2022). The Wsp chemosensory system modulates c-di-GMP-dependent biofilm formation by integrating DSF quorum sensing through the WspR-RpfG complex in Lysobacter. NPJ Biofilms and Microbiomes, 8, 97.

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Western Blot Analysis of His-Tagged Proteins

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Protein samples from separation of nickel column and nanoparticles were separated by SDS-PAGE and transferred to PVDF membrane. The membrane was subsequently blocked and incubated with primary antibody at 4 °C overnight. Anti-His-tag (1:2,000) was purchased from Abmart (Shanghai, China). The secondary antibody, anti-mouse (1:5000) was from Zhongshan Golden Bridge Biotechnology (Beijing, China). The blots were exposed to ECL Western Blotting Substrate (Vazyme, Nanjing, China).

Zhou Y., Yuan S., Liu Q., Yan D., Wang Y., Gao L., Han J, & Shi H. (2017). Synchronized purification and immobilization of his-tagged β-glucosidase via Fe3O4/PMG core/shell magnetic nanoparticles. Scientific Reports, 7, 41741.

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Validating Fusion Protein Expression in Exosomes

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To validate whether the fusion protein was successfully expressed in 293T cells and integrated into the exosome membranes, we performed western blot using anti-His antibody. Total protein of isolated exosomes or cells was extracted in RIPA Lysis Buffer (Solarbio, China) at 4 °C for 15 min. The protein concentration was determined using a BCA protein assay kit (Solarbio, China). About 30 µg of protein was separated on 10% SDS-PAGE gels, and then transferred to nitrocellulose membranes (Millipore). The membranes were blocked with 3% BSA, and then incubated with anti-His-tag (1:5000, Abmart, China, #M20001), and anti-β-actin (1:4000, Sigma, St Louis, MO, USA) at 4 °C overnight. Anti-CD63 (1:2000, Proteintech, Wuhan, Hubei, China, #25682-1-AP), anti-GM130 (1:2000, Proteintech, #11308-1-AP), anti-TSG101 (1:2000, Proteintech, #28283-1-AP) were also used to characterize the exosomes. anti-LPCAT1 (1:2000, Proteintech, #16112-1-AP), anti-EGFR (1:5000, Proteintech, #18986-1-AP). This procedure was followed by adding HRP-conjugated secondary antibody (1:10000, Cell Signaling Technology, Beverly, MA, USA) and ECL reagents (Solarbio, Beijing, China). The bands were visualized and recorded using the Tanon 5500 imaging system.

Jiang J., Lu Y., Chu J., Zhang X., Xu C., Liu S., Wan Z., Wang J., Zhang L., Liu K., Liu Z., Yang A., Ren X, & Zhang R. (2024). Anti-EGFR ScFv functionalized exosomes delivering LPCAT1 specific siRNAs for inhibition of lung cancer brain metastases. Journal of Nanobiotechnology, 22, 159.

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