THE™ His Tag Antibody, mAb, Mouse
THE™
His Tag Antibody, mAb, Mouse recognizes His tags localized at the C-terminal,
N-terminal, and internal region of fusion proteins.
| ￥2500 | |
| A00186-100 | |
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| ￥2500 | |
| A00186-100 | |
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| Specificity |
THE™
His Tag Antibody, mAb, Mouse recognizes His tags localized at the C-terminal,
N-terminal, and internal region of fusion proteins. |
| Host Species | Mouse |
| Immunogen |
A
synthetic peptide HHHHHH coupled to KLH |
| Conjugate | Unconjugated |
| Form | Lyophilized |
| Storage Buffer |
Lyophilized
with PBS, pH 7.4, containing 0.02% sodium azide. |
| Reconstitution |
Reconstitute
the lyophilized powder with sterile water (or equivalent) to a final
concentration of 0.5 mg/mL. |
| Storage Instructions |
The
lyophilized product remains stable up to 1 year at -20°C from date of receipt. The reconstituted antibody can be stored for
2-3 weeks at 2-8°C. For long term storage, aliquot and store at -20°C or below.
Avoid repeated freezing and thawing cycles. |
| Purification |
Purified
by Protein A affinity chromatography. |
| Isotype | Mouse IgG1,κ |
| Clonality | Monoclonal |
| Clone ID | 6G2A9 |
| Note |
GenScript
can offer this product according to your requirement, including product size,
buffer components, etc. |
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 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
»
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 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 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.
»
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. »
| Target Background |
Monoclonal
antibodies specific to six histidine tag 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 the His-tag. THE™ His Tag Antibody, mAb, Mouse (subtype IgG1) has very
high affinity towards the His-tag. 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. 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™ His Tag mAb recognizes His tags placed at N-terminal, C-terminal,
and internal regions of fusion proteins. THE™ His Tag mAb can be used in
Western blot analyses, Dot blot analyses, ELISA, immunofluorescent staining,
and flow cytometry of cultured cells. |
| Synonyms |
THE™
Anti-His mAb |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
