Mouse anti his tag antibody

Manufactured by Bio-Rad

Sourced in United States, United Kingdom

The Mouse-anti-His-tag antibody is a primary antibody specifically designed to detect the presence of the polyhistidine (His-tag) sequence in recombinant proteins. This antibody can be used in various immunoassay techniques to identify and purify His-tagged proteins.

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Lab products found in correlation

Hela cell, by American Type Culture Collection (1 mentions) Lipofectamine transfection reagent, by Thermo Fisher Scientific (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Rnx plus, by Sinaclon (1 mentions) Facs caliber, by BD (1 mentions) Hrp conjugated goat anti mouse igg, by Southern Biotech (1 mentions) His select nickel affinity gel, by Merck Group (1 mentions) Baculogold transfection kit, by BD (1 mentions) Elisa reader, by Dynex (1 mentions) O phenylenediamine substrate, by Merck Group (1 mentions) Goat anti mouse horseradish peroxidase hrp antibody, by Jackson ImmunoResearch (1 mentions)

Spelling variants of this product

Anti-His antibody (5 citations) Anti-His tag antibody (4 citations) Mouse anti-His (3 citations) Mouse anti-His antibody (2 citations) Mouse anti-6×His tag IgG1 antibody (2 citations)

4 protocols using mouse anti his tag antibody

In Vitro Expression of Mtb32C-HBHA Fusion Genes

Cited 2 times

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To confirm the in vitro expression of Mtb32C-HBHA fusion genes, the plasmid was transfected into HeLa cells (American Type Culture Collection Manassas, VA, USA) using lipofectamine transfection reagent according to the manufacturer’s instructions (Invitrogen, USA). Then, 72 hr after transfection, the cells were treated with 0.5 ml trypsin (Invitrogen, USA) and were incubated for 10 min. Cell suspension was harvested and total RNA was extracted using RNX-Plus (SinaClon, Iran), as described previously (16 (link), 17 (link)). Purified RNA was used for cDNA synthesis using cDNA synthesis kit (Pars Tous, Iran) and was amplified by PCR. To detect the presence of His-Tag marker in chimeric Mtb32C-HBHA protein (a marker of constructed rather than natural protein), Western blot method was performed using mouse anti-His Tag antibody as the primary antibody and peroxidase conjugated rabbit anti-mouse IgG as the secondary antibody (AbD SeroTec, USA).

Teimourpour R., peeridogaheh H., Teimourpour A., Arzanlou M, & Meshkat Z. (2017). A study on the immune response induced by a DNA vaccine encoding Mtb32C-HBHA antigen of Mycobacterium tuberculosis. Iranian Journal of Basic Medical Sciences, 20(10), 1119-1124.

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T Cell Bead-Mediated BsAb Binding Analysis

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T cells expanded by CD3/CD28 beads were reacted with BsAbs (1µg per 10⁶ cells) for 30 min on ice. After washing, the T cells were reacted with a mouse-anti-His-tag antibody (AbD Serotec) (0.5 µg per 10⁶) on ice for 30 min. A goat anti-mouse Alexa-647 conjugated antibody was finally added to the washed T cells. After 20 min of reaction, T cells were washed and analyzed by flow cytometry on the FACS Caliber (BD Biosciences, San Jose, CA)

Cheng M., Ahmed M., Xu H, & Cheung N.K. (2014). Structural design of disialoganglioside GD2 and CD3-bispecific antibodies to redirect T cells for tumor therapy. International journal of cancer. Journal international du cancer, 136(2), 476-486.

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Production and Purification of Recombinant rbFcRn

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Recombinant baculoviruses carrying the cDNAs of the soluble rbFcRn α-chain and rbβ2m were generated by using the BaculoGold Transfection Kit (BD Biosciences Pharmingen, Franklin Lakes, NJ, USA) according to the manufacturer’s instructions. Then, Sf9 cells were cultured and infected by recombinant viruses. Recombinant rbFcRn was purified from supernatants of virus infected cells by using Ni-NTA chromatography (His-Select Nickel Affinity Gel, Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer’s protocol. Expression level and purity of the recombinant protein was confirmed by 15% SDS-PAGE followed by either Coomassie staining or Western blotting. Mouse anti-His tag antibody (AbD Serotec, Kidlington, UK) as the primary (500x) and HRP-conjugated goat anti-mouse IgG (Southern Biotech, Birmingham, AL, USA) as the secondary antibody (10.000x) were used for immunodetection. The yield of the rbFcRn expression was approximately 5 mg/l (5 mg purified protein per 1 liter cell culture supernatant).

Szikora B., Hiripi L., Bender B., Kacskovics I, & Iliás A. (2017). Characterization of the interactions of rabbit neonatal Fc receptor (FcRn) with rabbit and human IgG isotypes. PLoS ONE, 12(9), e0185662.

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Quantification of Bispecific Antibody Binding

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GD2 was first coated at 1 µg/ml per well in 90% ethanol on vinyl 96-well plates at room temperature (RT) overnight. For CD3 binding, 5000/per well of Jurkat cells (100 µl in PBS) were used to coat 96-well plates overnight. The next day, after blocking the plates with 0.5% BSA (150 µl/well), serial dilutions of the BsAb antibodies were added to the microtiter wells and incubated at RT for 2 h. Plates were washed with PBS, a mouse-anti-His-tag antibody (AbD Serotec) (1:1000 dilutions in 0.5% BSA; 100 µl/well) was added and incubated for another hour. After washing, the wells were reacted with a goat-anti-mouse-horse radish peroxidase (HRP) antibody (1:3000 dilutions in 0.5% BSA; 100 µl/well) (Jackson ImmunoResearch) and incubated at RT for 1h. Color was developed using the o-phenylenediamine substrate (Sigma) and the ODs measured at 490 nm using ELISA reader (Dynex Technologies).

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