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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

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.
A00186S
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Overview
Specificity THETM 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

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

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 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.
Synonyms THETM Anti-His mAb;

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