WB
Western blot analysis of 6X His tag on different lysates with Rabbit anti-6X His tag antibody at 1/5,000 dilution. Lane 1: 293T transfected with His-tagged empty control cell lysate, Lane 2: 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate, Lane 3: 293T transfected with His-tagged Histone H3.1 (C-terminal) cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 2 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded HeLa transfected with His-tagged Histon H3.1 (C-terminal) cells with Rabbit anti-6X His tag antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling 6X His tag with Rabbit anti-6X His tag antibody at 1/5,000 dilution. HeLa cells, transfected with His-tagged Histone H3.1 (C-terminal) or ACAT2 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-6X His tag antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa transfected with His-tagged ACAT2 (N-terminal) cells labeling 6X His tag. Cells were fixed and permeabilized. Then stained with the primary antibody (1/200) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
6X His tag was immunoprecipitated from 0.2 mg 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate with Rabbit anti-6X His tag antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-6X His tag antibody at 1/500 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate (input), Lane 2: Rabbit anti-6X His tag antibody IP in 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-6X His tag antibody in 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 8 seconds.| Product Name | 6X His tag Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide immune sequence is HHHHHHHHC. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:5000 ICC/IF:1:5000 FC:1:200-1:500 IP:1-2μg/sample |
|
| Species Reactivity | All |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | 6 His epitope tag antibody Hexa His tag antibody HHHHHH epitope tag antibody HHHHHH tag antibody His tag antibody Polyhistidine Tag antibody |
|---|---|
| Function | His-Tag Antibody detects recombinant proteins containing the 6xHis epitope tag. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells. |
WB
Western blot analysis of 6X His tag on different lysates with Rabbit anti-6X His tag antibody at 1/5,000 dilution. Lane 1: 293T transfected with His-tagged empty control cell lysate, Lane 2: 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate, Lane 3: 293T transfected with His-tagged Histone H3.1 (C-terminal) cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 2 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded HeLa transfected with His-tagged Histon H3.1 (C-terminal) cells with Rabbit anti-6X His tag antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling 6X His tag with Rabbit anti-6X His tag antibody at 1/5,000 dilution. HeLa cells, transfected with His-tagged Histone H3.1 (C-terminal) or ACAT2 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-6X His tag antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa transfected with His-tagged ACAT2 (N-terminal) cells labeling 6X His tag. Cells were fixed and permeabilized. Then stained with the primary antibody (1/200) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
6X His tag was immunoprecipitated from 0.2 mg 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate with Rabbit anti-6X His tag antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-6X His tag antibody at 1/500 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate (input), Lane 2: Rabbit anti-6X His tag antibody IP in 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-6X His tag antibody in 293T transfected with His-tagged ACAT2 (N-terminal) cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 8 seconds.| Application Notes | WB:1:5000 IHC:1:5000 ICC/IF:1:5000 FC:1:200-1:500 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
