THE™ His Tag Antibody, mAb, Mouse

Monoclonal antibodies specific to six histidine tags can greatly improve the effectiveness of several different kinds of immunoassays, helping researchers identify, detect, and purify polyhistidine fusion proteins in bacteria, insect cells, and mammalian cells. However, since 6XHis-tag is poorly immunogenic, it needs to be conjugated to KLH or some other carrier as an immunogen. After hundreds of selection cycles, researchers at GenScript successfully isolated an antibody against His-tag. THETM His Tag Antibody, mAb, Mouse (subtype IgG1) has very high affinity. Tests performed at GenScript show that the antibody can also recognize 4xHis- and 5xHis-tags. This means that even if the 6xHis-tag is only partially exposed, it will still be recognized and bound by this antibody. THETM His Tag mAb is produced from mice ascites and purified by protein A affinity column. This antibody recognizes native as well as denatured forms of synthetic polyhistidine and polyhistidine-tagged fusion proteins. The product reacts with fusion proteins expressed in bacteria, insect cells, and mammalian cells. THETM His Tag mAb recognizes His tags placed at N-terminal, C-terminal, and internal regions of fusion proteins. THETM His Tag mAb can be used in Western blot analyses, Dot blot analyses, ELISA, immunofluorescent staining, and flow cytometry of cultured cells.

Price	$0.00
Cat. No.	A00186S
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Overview Properties Background

Overview

Specificity	THE^TM His Tag Antibody, mAb, Mouse recognizes C-terminal, N-terminal, and internal His tagged fusion proteins.
Host Species	Mouse
Immunogen	A synthetic peptide HHHHHH coupled to KLH

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Properties

Concentration	0.5 mg/ml, lyophilized with PBS, pH 7.4, containing 0.02% sodium azide
Reconstitution	Reconstitute the lyophilized antibody with deionized water (or equivalent) to a final concentration of 0.5 mg/ml.
Purification	Protein A affinity column
Clone ID	6G2A9

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Figure 1. Comparative His-tag detection by Western blot was performed using THE™ His Antibody, mAb, Mouse (A: GenScript, A00186, at 0.1 µg/mL concentration) and Mouse Anti-His mAb (B: Competitor A, at 0.1 µg/mL concentration).
Both antibodies were used to probe the same sample containing overexpressed His-tagged fusion protein. »

Figure 2. Comparative His-tag detection by Western blot was performed using THE™ His Antibody, mAb, Mouse (A: GenScript, A00186, at 1 µg/mL concentration) and Mouse Anti-His mAb (B: Competitor B, at 1 µg/mL concentration).
Both antibodies were used to probe the same cell lysates containing His-tagged fusion protein.
The signal was developed with Goat Anti-Mouse IgG (H&L) [HRP] Polyclonal Antibody.
Researchers can also use MonoRab™ Anti-Mouse IgG (H&L) (76F10), mAb, Rabbit (GenScript, V90301) as a secondary antibody. »

Figure 3. Comparative His-tag detection by Dot blot was performed using THE™ His Antibody, mAb, Mouse (A: GenScript, A00186, at 1 µg/mL concentration) and two Mouse Anti-His mAbs (B: Competitor Q#1, at 1 µg/mL concentration; C: Competitor Q#2, at 1 µg/mL concentration).
All three antibodies were used to probe the same samples containing His-tagged fusion protein.
The signal was developed with Goat Anti-Mouse IgG (H&L) [HRP] Polyclonal Antibody.
Researchers can also use MonoRab™ Anti-Mouse IgG (H&L) (76F10), mAb, Rabbit (GenScript, V90301) as a secondary antibody. »

Figure 4. Lot-to-lot consistency of antibody performance was analyzed for 4 batches (Batched 1#, 2#, 3# and 4#) of THE™ His Antibody, mAb, Mouse (GenScript, A00186, 1 µg/mL) by Western blot.
The results show that the antibody-generated signal remains consistent from Lot-to-Lot.
Antibodies from all four lots were used to probe samples of the same His-tagged fusion protein.
The signal was developed with IRDye™ 800 Conjugated Goat Anti-Mouse IgG. »

Figure 5. Detection of His-tag in CHO cells transfected with His-tagged protein (Green), and non-transfected CHO cells (Black) by flow cytometry using THE™ His Tag Antibody, mAb, Mouse (GenScript, A00186).
The signal was developed with FITC conjugated Goat Anti-Mouse IgG. »

Figure 6. Detection of N- or C-terminal His tags in recombinant fusion proteins were analyzed by Western blot using THE™ His Antibody, mAb, Mouse (GenScript, A00186, at 1 µg/mL concentration).
Lane 1: N-terminal His-tagged fusion protein
Lane 2: C-terminal His-tagged fusion protein
The signal was developed with Goat Anti-Mouse IgG (H&L) [HRP] Polyclonal Antibody.
Researchers can also use MonoRab™ Anti-Mouse IgG (H&L) (76F10), mAb, Rabbit (GenScript, V90301) as a secondary antibody.
The results show that THE™ His Antibody, mAb, Mouse (GenScript, A00186) can recognize N-terminal and C-terminal His-tagged proteins. »

Figure 7. Immunoprecipitates from cell lysates containing His-tagged fusion protein were analyzed by Western blot using THE™ His Antibody, mAb, Mouse (GenScript, A00186).
Lane 1. Positive control containing His-tagged fusion protein
Lane 2. Negative control – IP with isotype control antibody (A01007)
Lane 3. Immunoprecipitation with THE™ His Tag Antibody, mAb, Mouse (A00186) »

Figure 8. His-tag was detected in Multiple Tag Cell Lysate (GenScript, M0100) by Western blot using THE™ His Antibody, mAb, Mouse (GenScript, A00186, 1 µg/mL).
The signal was developed with Goat Anti-Mouse IgG (H&L) [HRP] Polyclonal Antibody.
Researchers can also use MonoRab™ Anti-Mouse IgG (H&L) (76F10), mAb, Rabbit (GenScript, V90301) as a secondary antibody.
His-tagged fusion protein:
Predicted MW: 52 kDa
Observed MW: 52 kDa »

BLI (Biolayer interferometry) binding affinity measurements of THE™ His Tag Antibody, mAb, Mouse to different His-tagged fusion proteins.
The results show that THE™ His Tag Antibody, mAb, Mouse (GenScript, A00186) is able to recognise the His tag at different positions in the protein. »

Background

Target Background

Monoclonal antibodies specific to six histidine tags can greatly improve the effectiveness of several different kinds of immunoassays, helping researchers identify, detect, and purify polyhistidine fusion proteins in bacteria, insect cells, and mammalian cells. However, since 6XHis-tag is poorly immunogenic, it needs to be conjugated to KLH or some other carrier as an immunogen. After hundreds of selection cycles, researchers at GenScript successfully isolated an antibody against His-tag.
THE^TM His Tag Antibody, mAb, Mouse (subtype IgG1) has very high affinity. Tests performed at GenScript show that the antibody can also recognize 4xHis- and 5xHis-tags. This means that even if the 6xHis-tag is only partially exposed, it will still be recognized and bound by this antibody.
THE^TM His Tag mAb is produced from mice ascites and purified by protein A affinity column. This antibody recognizes native as well as denatured forms of synthetic polyhistidine and polyhistidine-tagged fusion proteins. The product reacts with fusion proteins expressed in bacteria, insect cells, and mammalian cells. THE^TM His Tag mAb recognizes His tags placed at N-terminal, C-terminal, and internal regions of fusion proteins.
THE^TM His Tag mAb can be used in Western blot analyses, Dot blot analyses, ELISA, immunofluorescent staining, and flow cytometry of cultured cells.

Synonyms

THE^TM Anti-His mAb;

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Citations

1. Greeshma Jain, et al. Inhibitor-based modulation of huntingtin aggregation mechanisms mitigates ﬁbrilinduced cellular stress. Nat Commun. (2025-04)