Catalog Products » Antibodies » Epitope Tag Antibodies » THE™ His Tag Antibody, mAb, Mouse
THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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.

THE™ His Tag Antibody, MAb, Mouse

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)

THE™ His Tag Antibody, MAb, Mouse

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

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. 
A00186
$385.00

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Overview
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

Applications
Working concentrations for specific applications should be determined by the investigator. The appropriate concentrations may be affected by secondary antibody affinity, antigen concentration, the sensitivity of the method of detection, temperature, the length of the incubations, and other factors. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product.
Application Recommended Usage
ELISA 0.05-0.2 µg/ml
Western Blot 0.1-0.2 µg/ml
Immunoprecipitation (IP) 1 µg/ml
Immunocytochemistry/Immunofluorescence (ICC/IF) 1 µg/ml
Flow Cytometry 1 µg/ml
Time-resolved fluorescence energy transfer (TR-FRET) User-optimized

Properties
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.

Examples
  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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.

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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)

  • THE™ His Tag Antibody, MAb, Mouse
  • THE™ His Tag Antibody, MAb, Mouse

    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


Background
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.


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