THE™ His Tag Antibody (Mouse)
A Gold Standard for assay development and quality control
THE™ His tag antibody recognizes 6×His-, 5×His-, and 4×His-tags in the N-terminal, C-terminal, or internal region of fusion proteins. It is widely used to detect the existence of His-tagged proteins in various applications including HTRF assays for drug discovery, FACS, ICC/IF, ChIP, WB, affinity purification, capture, ELISA, and more. Its affinity is in the low nanomolar range, which makes THE™ His tag antibody the most sensitive on the market. THE™ His tag antibody has been on the market for 10 years, and is widely accepted by scientists in industry as a standard reagent for internal QC and assay development.
- High Specificity - No non-specific binding
- Superior Sensitivity - The most sensitive His Tag mAb on the market
- Excellent Affinity- Marked lower dissociation, constant Kd
- Improved Reactivity- All C-terminal, internal, and N-terminal His Tags can be identified
- Enhanced Binding - Detects even exposed 6xHis-Tags, 5xHis-Tags, and 4xHis-Tags
- Long-term Stability - Lot-to-lot consistency and stable lyophilized form for at least two years
1. Western Blot of 6xHis-tag Fusion Protein using THE™ His Tag Antiboy (A00186)
Figure 1: Sensitivity comparison of 6xHis-tag Fusion Protein by Western blot using His Antibody, mAb, Mouse from GenScript (A00186, 0.1 μg/ml) and Qiagen (34698 , 0.1 μg/ml).
Figure 2: Western blot analysis of N-terminal and C-terminal 6xHis-tag Fusion Protein using THE™ His Antibody, mAb, Mouse (A00186, 1 μg/ml).
2. Dot Blot Detection of 5xHis-GST in Whole Cell Lysates using THE™ His Tag Antibody (A00186)
Figure 3: Sensitivity comparison of 5xHis-GST in Whole Cell Lysates by Dot blot using His Antibody, mAb, Mouse from GenScript(A: A00186, 1 μg/ml) and Qiagen (B: 34698, 1 μg/ml and C:34660, 1 μg/ml).
3. Western Blot Detection of 4xHis-GST in Whole Cell Lysates using THE™ His Tag Antibody (A00186)
Figure 4: Sensitivity comparison of 4xHis-GST in Whole Cell Lysates by Western blot using His Antibody, mAb, Mouse from GenScript (A00186, 1 μg/ml) and Qiagen (34698 , 1 μg/ml).
4. Western blot analysis of immunoprecipitates using THE™ His Tag Antibody (A00186)
Figure 5: Western blot analysis of immunoprecipitates from cell lysates containing His fusion protein using THE™ His Antibody, mAb, Mouse (A00186).
5. Lot-to-lot consietency using Western Blotting
Figure 6: Consistency analysis of THE™ His Antibody, mAb, Mouse (GenScript, A00186, 1 μg/ml) with His-tagged fusion protein by Western blot.
6. Immunofluorescent staining using THE™ His Tag Antibody (A00186)
Figure 7: Immunocytochemistry/Immunofluorescence analysis of HEK293 cells (A) and HEK293 cells transfected with His tagged fusion gene (B) using THE™ His Antibody, mAb, Mouse (A00186).
7. Flow cytometric analysis using THE™ His Tag Antibody (A00186)
Figure 8: Flow cytometric analysis of HEK293 cells (A) and HEK293 cells transfected with His tagged fusion gene (B) using THE™ His Antibody, mAb, Mouse (A00186).
- Will using a brighter fluorophore allow better detection of a molecule on a cell that is in low abundance?
The brightness of fluorophore and the antibody's sensitivity are two factors that influence detection sensitivity. GenScript's THE series antibodies have the best sensitivity on the market. The brighter the fluorophore is, the better the overall detection.
GenScript's IFluor antibody series are newly developed fluorescent compounds conjugated to our THE series antibodies. Compared with traditional fluorescent dyes, the IFluor series are brighter and have better stability.
- Which antibody should I use to detect a very small amount of His protein in a Flow Cytometry assay?
We have four types of fluorophore conjugated His-tag antibodies for FACS analysis, including A01620(FITC), A01800(iFluor488), A01801(iFluor555) and A01802(iFluor 647).
The iFluor series are recommended for FACS analysis because they have a higher sensitivity than FITC conjugated antibody. The following is the wavelength range of GenScript's iFluor antibodies:
Cat. No. Conjugate Ex (nm) Em (nm)A01800iFluor 488491514A01801iFluor 555559569A01802iFluor 647654674
According to our FACS analysis results, the sensitivity of A01802 (THE™ His Tag Antibody [iFluor 647], mAb, Mouse) is slightly higher than the A01800 (THE™ His Tag Antibody [iFluor 488], mAb, Mouse). If the His Tag number is low, A01802 is recommended.
- For A00186 (THE™ His Tag Antibody, mAb, Mouse), what is the working concentration range for Immunofluorescence?
For A00186, a starting point of 1 µg/ml is recommended.
- We are successfully using your ELISA kit L00436 (His Tag ELISA Detection Kit), and we would like to use the same anti-His monoclonal antibody for western blot. What is the Catalog No. for the mAb used in the L00436 ELISA kit? What is its recommended concentration for WB?
The catalog No. is A00186 for the anti-His mAb in the L00436 ELISA Kit. The recommended concentration for WB is 0.1-0.2 µg/ml.
- I need a rabbit anti-His tag antibody which can recognize a His-tag fused in the internal site of a protein. I want to perform Flow Cytometry with this antibody. Which antibody should I use?
A. Catalog No. A00174 is a rabbit anti-His tag antibody. It has been tested for internal His-tag recognition, but not for Flow Cytometry.
For Flow Cytometry, the following antibodies are recommended. All of them can recognize His-tag fused on the internal site of a protein.