| Description | This product is a monoclonal antibody derived from Mouse (Mus musculus), which can specifically recognize Polyhistidine-tag. The antibody is expressed with mammalian cell transient expression system, serum-free and purified by affinity chromatography. The purity and integrity are tested via SDS-PAGE and SEC-HPLC analysis. Given an antigen, additional QC measures are also desired such as affinity testing and binding validation. Specifically, the antibody is provided in multiple formats for in vivo and in vitro assays. The Invivo version features greater than 95% purity, ultra-low endotoxin levels (<1 EU/mg or 0.1 EU/mg), and is preservative, stabilizer, and carrier protein-free. |
| Clonality | Monoclonal |
| Host Species | Mouse |
| Isotype | IgG |
| Expression Species | HEK293F or CHO |
| Conjugation | None |
| Purity | >95%, determined by SDS-PAGE and/or SEC-HPLC |
| Endotoxin | <1 EU/mg, determined by LAL method |
| Purification | Protein A affinity purified |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4⁰C within a week. For longer storage, aliquot and store at -20⁰C. |
| Application | DB; WB; ELISA; FuncS |
| Application Notes | The antibody is recommended for detection of His Tag by WB, ELISA, FuncS assays. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
| Target | His Tag |
| Alternative Name | hexa histidine-tag; 6xHis-tag; His6 tag; His Tag; Polyhistidine-tag |
| Research Area | Cell Biology |
Figure 1 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in WB
Western blot analysis of V3S-0522-YC7376 was performed by loading a protein with His tag onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with V3S-0522-YC7376 and HRP goat anti-mouse IgG (H+L) as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.6 μg)
Figure 1 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in WB
Figure 2 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in DB
Dot Blot analysis of V3S-0522-YC7376 was performed by coating with a protein with His Tag.
V3S-0522-YC7376 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-mouse IgG (H+L) as a secondary antibody (1: 2000)
NC: Negative control PC: Positive control
Figure 2 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in DB
Figure 3 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in ELISA
ELISA analysis of V3S-0522-YC7376 was performed by coating with a protein with His Tag. Then blocked with BSA and incubated with V3S-0522-YC7376. The HRP-conjugated goat anti-Mouse IgG (H+L) as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 3 Anti-His Tag Monoclonal Antibody (V3S-0522-YC7376) in ELISA
Figure 4 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in SDS-PAGE.
SDS-PAGE analysis of V3S-0522-YC7376 Fab fragment in non-reduced (Lane 1, 1.5 μg) and β-mercaptoethanol-reduced (Lane 2, 1.5 μg) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrated as about 25 kDa. And non-reduced protein migrated about 35-48 kDa.
Figure 4 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in SDS-PAGE.
Figure 5 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in Western Blot.
Western blot analysis of V3S-0522-YC7376 Fab fragment was performed by loading a protein with His tag onto a 12% Tris-HCl polyacrylamide gel. proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with the Fab and HRP goat anti-mouse IgG (H+L) as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.1 μg)
Lane 2: Reduced antigen (0.3 μg)
Lane 3: Reduced antigen (0.6 μg)
Figure 5 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in Western Blot.
Figure 6 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in ELISA.
ELISA analysis of V3S-0522-YC7376 Fab fragment was performed by coating with a protein with His Tag. Then blocked with BSA and incubated with the Fab. The HRP-conjugated goat anti-mouse IgG (H+L) as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 6 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in ELISA.
Figure 7 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in Dot Blot.
Dot blot analysis of V3S-0522-YC7376 Fab fragment was performed by coating with a protein with His Tag.
V3S-0522-YC7376 Fab fragment incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-mouse IgG (H+L) as a secondary antibody (1: 5000)
NC: Negative control PC: Positive control
Figure 7 Recombinant Anti-His Tag Antibody (V3S-0522-YC7376) Fab Fragment in Dot Blot.
