His tag monoclonal antibody his h8

Manufactured by Thermo Fisher Scientific

The 6x-His-Tag monoclonal antibody (His.H8) is a laboratory reagent used for the detection and purification of recombinant proteins with a 6x-histidine (6x-His) tag. This antibody is specific to the 6x-His tag, allowing it to bind and identify proteins that have been engineered to contain this tag.

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Lab products found in correlation

Goat anti mouse igg labeled with horse radish peroxidase, by Thermo Fisher Scientific (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions)

Spelling variants of this product

6x-His Tag monoclonal antibody (16 citations) His-tag antibody (15 citations) His-tag (7 citations) HIS.H8 (4 citations)

2 protocols using his tag monoclonal antibody his h8

Expression and Detection of Camel IFNε in E. coli

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The expression of the camel recombinant IFNε gene in E. coli was checked by performed a 12% SDS-PAGE according to Laemmli, 1970 [32 (link)]. After electrophoresis, the gel was stained with Coomassie Brilliant Blue R-250 followed by de-staining in a solution of 10% (v/v) methanol and 10% (v/v) acetic acid. Recombinant C. dromedarius IFNε protein was detected by wesetrn blotting using 6x-His-Tag monoclonal antibody (His.H8, Cat# MA1-21315, ThermoFischer Scientific) at a 1:1000 dilution according to Towbin et al. [33 (link)]. Secondary antibody used was goat anti-mouse IgG labeled with horse radish peroxidase (Invitrogen Cat# G-21040) at a 1:2000 dilution. The membrane was developed using chromogenic substrate 3,3’,5,5’-Tetramethyl benzidine (TMB) liquid substrate system (Sigma-Aldrich, Cat# T0565).

Abdel-Fattah M., Saeed H., El-Shennawy L., Shalaby M., Embaby A., Ataya F., Mahmoud H, & Hussein A. (2019). The Arabian camel, Camelus dromedarius interferon epsilon: Functional expression, in vitro refolding, purification and cytotoxicity on breast cancer cell lines. PLoS ONE, 14(9), e0213880.

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Quantification of Recombinant Procaspase-8

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The concentration of recombinant procaspase-8 proteins was calculated by NanoDrop™ and the final protein concentrations adjusted to 500 nM in clarified HEK 293T cellular lysates. The procaspase-containing lysates were then resolved by SDS–PAGE and transferred to PVDF membranes, blocked with 5% milk in TBST and probed with 6x-His Tag Monoclonal Antibody (HIS.H8) (ThermoFisher MA1–21315, 1:3000). Blots were incubated with primary antibodies overnight at 4 °C with rocking and were then washed (3×5min, TBST) and incubated with secondary antibodies (LICOR, IRDye 800LT goat anti-mouse, 1:10,000) for 1h at ambient temperature. Blots were further washed (3× 5min, TBST) and visualized on a BioRad ChemiDoc™ MP Imaging System.

Xu J.H., Eberhardt J., Hill-Payne B., González-Páez G.E., Castellón J.O., Cravatt B.F., Forli S., Wolan D.W, & Backus K.M. (2020). Integrative x-ray structure and molecular modeling for the rationalization of procaspase-8 inhibitor potency and selectivity. ACS chemical biology, 15(2), 575-586.

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