Anti-6X His tag® antibody [HIS.H8] is a mouse monoclonal antibody that is used to detect 6X His tag® in Dot, ELISA, ICC/IF, IP, Western blot.
- Cited in over 350 publications
- Binds to his-tagged proteins
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS
IP | ELISA | Dot | WB | ICC/IF | |
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Species Tag | Dilution info 1/200 | Notes - |
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Species Tag | Dilution info Use at an assay dependent concentration. | Notes - |
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Species Tag | Dilution info Use at an assay dependent concentration. | Notes - |
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Species Tag | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
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Species Tag | Dilution info 1/200 | Notes - |
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6 His epitope tag, HHHHHH epitope tag, HHHHHH tag, Hexa His tag, Polyhistidine Tag
Anti-6X His tag® antibody [HIS.H8] is a mouse monoclonal antibody that is used to detect 6X His tag® in Dot, ELISA, ICC/IF, IP, Western blot.
- Cited in over 350 publications
- Binds to his-tagged proteins
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS
Recognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells.
ab18184 reacts to recombinant proteins containing the 6X His tag® or 10X His tag® fused to either the amino or carboxy terminus.
Purified from TCS by Protein A affinity chromatography
Anti-6X His tag® antibody [HIS.H8] (ab18184) is a mouse monoclonal antibody and is validated for use in Dot, ELISA, ICC/IF, IP and WB.
Anti-6X His tag® antibody [HIS.H8] (ab18184) was first used in a scientific publication in 2004 and has been cited over 356 times in peer reviewed journals. It's performance in Western blot in human and mouse samples is trusted by the scientific community.
Abcam's high quality validation processes ensure Anti-6X His tag® antibody [HIS.H8] (ab18184) has high sensitivity and specificity.
Anti-6X His tag® antibody [HIS.H8] (ab18184) has 31 independent reviews from customers.
6X His tag® antibodies are used to visualize proteins labelled with this tag in a variety of applications (for example imaging and Flow cytometry). To enable specific detection of your tagged protein, Anti-6X His tag® antibody [HIS.H8] (ab18184) has been validated in Dot, ELISA, ICC/IF, IP and WB.
Anti-6X His tag® antibody [HIS.H8] (ab18184) specifically detects 6X His tag® (UniProt ID: P26367; Molecular weight: 47kDa) and is sold in 100 ug selling sizes.
Antibody clone HIS.H8 is also available pre-conjugated to a variety of labels for your convenience - Biotin (Biotin Anti-6X His tag® antibody [HIS.H8] ab173828).
Top cited 6X His tag antibody in the market with >450 citations. The 6X His tag, also known as a polyhistidine tag, is widely used for the purification of His-tagged proteins. Utilizing Ni-NTA chromatography, this tag facilitates efficient protein purification, ensuring high yield and purity. It is also highly relevant for detecting 6X His tags in multiple proteomic applications such as western blotting (WB), Immunoprecipitation (IP) and immunocytochemistry/immunofluorescence (ICC/IF) applications.
HIS-TAG® is a trademark of EMD Biosciences, Inc.
The 6X His tag is a sequence commonly used for protein purification detection and immobilization. It consists of six histidine residues that bind to metal ions like nickel or cobalt. This makes it a highly efficient tag for affinity chromatography. His tag antibodies such as anti-His HRP facilitate the detection of His-tagged proteins in various assays. Being a small tag with a typical molecular weight of about 0.84 kDa it minimally interferes with protein function. Many proteins expressed in E. coli and other expression systems carry the 6X His tag for ease of purification and analysis.
Proteins tagged with the 6X His tag often play roles in studies involving recombinant proteins. The tag itself does not alter the biological function of the target protein. It is not part of any natural protein complex but aids in the study of such complexes. Researchers use His tag antibodies like anti-His to specifically target and study these tagged proteins without cross-reactivity to other proteins.
The incorporation of the 6X His tag into proteins allows them to be studied more efficiently in various biological pathways. It can be part of pathways like signal transduction or metabolic pathways. When tagged proteins interact with other molecules such as kinases or receptors researchers can track these interactions using the tag and anti-Histidine antibodies. This technique informs on proteins that are hard to purify due to low abundance or instability.
The 6X His tag itself does not have direct implications. However proteins of interest in disease models often carry the tag which allows researchers to purify and study them in detail. For example cancer-related proteins may be expressed with a His tag to understand their role in tumorigenesis. Similarly proteins involved in metabolic disorders could be tagged which helps in analyzing specific interactions or functions disrupted in diseases.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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Terms & Conditions.
All lanes: Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/1000 dilution
Lane 1: 6x His-tagged protein expressed in BL21 (pET15b) at 1 µg
Lane 2: 6x His-tagged protein expressed in BL21 (pET15b) at 0.1 µg
All lanes: IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution
Performed under reducing conditions.
Exposure time: 5min
Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 at 1/100 dilution.
Lane 1 : MW Ladder (from top 120, 85, 50, 34, 25, 19 kDa)
Lanes 2-5 : Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 from mixture of ~50 ng purified His/FLAG tagged PDCD5 and 15 μL RRL in 300 μL buffer.
Lanes 6-9 : Purified PDCD5 from E. coli with ~60-70 ng protein.
The lowest band in each lane is ~ 20 kDa and is PDCD5. The upper bands in lanes 2-5 are the heavy chain (~50 kDa, runs as doublet) and light chain (~25 or 26 kDa).
Western Blot was performed with a mouse anti-FLAG primary antibody at 1/2000 dilution. An IRDye®800CW-conjugated goat anti-mouse IgG was used as a secondary antibody.
All lanes: Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)
Blocked with 5% milk for 30 minutes at 25°C.
Incubated with the primary antibody for 16 hours at 4°C in 1X PBS/Tween, 5% milk.
All lanes: Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution
Lane 1: ARPE-19 whole cell lysate - non-transfected at 20 µg
Lane 2: ARPE-19 whole cell lysate - transfected with rRab27a-His at 20 µg
All lanes: HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 5min
ab18184 at a 1/1000 dilution staining 6X His tag in transfected human HEK293 cells by Immunocytochemistry/ Immunofluorescence incubated for 12 hours at 22°C. PFA fixed. Permeabilized using 0.1% Triton X-100 in PBS. Blocked using 3% BSA for 1 hour at 22°C. Secondary used at a 1/1000 dilution polyclonal Goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 488 (green). DAPI staining (blue).
LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).
Anti-SARS-CoV-2 nsp13 antibody [EPR24838-18] ab286909 was shown to bind specifically to SARS-CoV-2 nsp13 protein in western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
Blocking buffer: 5% milk in TBS-0.1% Tween® 20 (TBS-T)
Lanes 1 - 4: Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/1000 dilution
Lanes 5 - 8: Western blot - Anti-SARS-CoV-2 nsp13 antibody [EPR24838-18] (Anti-SARS-CoV-2 nsp13 antibody [EPR24838-18] ab286909) at 1/1000 dilution
Lanes 1 and 5: Recombinant SARS-CoV-2 nsp13 protein (His-tagged) at 0.5 µg
Lanes 2 and 6: Recombinant SARS-CoV-2 nsp13 protein (His-tagged) at 0.2 µg
Lanes 3 and 7: Recombinant SARS-CoV2 nsp1 protein (His tagged) at 0.5 µg
Lanes 4 and 8: Recombinant SARS-CoV2 nsp1 protein (His tagged) at 0.2 µg
Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Lanes 5 - 8: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
In Western blot, Anti-SARS-CoV-2 nsp14 antibody [EPR24839-87] ab286912 was shown to bind specifically to NSP14. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.
All lanes: Western blot - Anti-SARS-CoV-2 nsp14 antibody [EPR24839-87] (Anti-SARS-CoV-2 nsp14 antibody [EPR24839-87] ab286912) at 1/1000 dilution
Lane 1: Recombinant SARS-CoV-2 NSP14 protein - Active (His-Tagged) at 0.5 µg with 3 % milk in TBS-0.1 % Tween® 20 (TBS-T)
Lane 3: Recombinant SARS-CoV-2 NSP14 (His-tagged) at 0.5 µg with 3 % milk in TBS-0.1 % Tween® 20 (TBS-T)
Lane 5: SARS-CoV-2 NSP1 protein (His-tagged) at 0.5 µg with 3 % milk in TBS-0.1 % Tween® 20 (TBS-T)
Lane 7: Recombinant SARS-COV-2 NSP2 protein (His-tagged) at 0.5 µg with 3 % milk in TBS-0.1 % Tween® 20 (TBS-T)
Lane 9: Recombinant SARS-CoV-2 NSP9 Protein (His-tagged) at 0.5 µg with 3 % milk in TBS-0.1 % Tween® 20 (TBS-T)
Lanes 1 - 9: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 9: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 64 kDa
Lane 1: Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active, His tagged) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 ug
Lane 2: Recombinant SARS-COV-2 NSP2 protein (His tagged) at 0.2 ug
False colour image of Western blot: Rabbit monoclonal [EPR24852-116] to SARS-CoV-2 Spike Glycoprotein S1 Chicken IgY (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Chicken IgY (Chimeric) ab323001) staining at 1/500 dilution, shown in black; Mouse Monoclonal 6X His tag® (ab18184) loading control staining at 1/1000 dilution, shown in red. In Western blot, Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Chicken IgY (Chimeric) ab323001 was shown to bind specifically to SARS-CoV-2 spike glycoprotein S1. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Chicken IgY H&L (HRP) preadsorbed (Goat Anti-Chicken IgY H&L (HRP) preadsorbed ab7118) at 1:10000 and Goat anti-Mouse IgG H&L 680RD at 1:20000.
All lanes: Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Chicken IgY (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Chicken IgY (Chimeric) ab323001) at 1/500 dilution
Lane 1: Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 µg
Lane 2: Recombinant SARS-COV-2 NSP2 protein (His tagged) at 0.2 µg
Lanes 1 - 2: Western blot - Goat Anti-Chicken IgY H&L (HRP) preadsorbed (Goat Anti-Chicken IgY H&L (HRP) preadsorbed ab7118) at 1/10000 dilution
Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 135 kDa
Exposure time: 4min
Lane 1: Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active, His tagged) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 ug
Lane 2: Recombinant SARS-COV-2 NSP2 protein (His tagged) at 0.2 ug
Performed under reducing conditions.
Observed band size: 135 kDa.
False colour image of Western blot: Rabbit monoclonal [EPR24852-116] to SARS-CoV-2 Spike Glycoprotein S1 - Human IgG1 (Chimeric), Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Human IgG1 (Chimeric) ab323000 staining at 1/500 dilution, shown in black; Mouse Monoclonal 6X His tag® (ab18184) loading control staining at 1/1000 dilution, shown in red. In Western blot, Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Human IgG1 (Chimeric) ab323000 was shown to bind specifically to SARS-CoV-2 spike glycoprotein S1. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Human IgG Fc (HRP) preadsorbed (Goat Anti-Human IgG Fc (HRP) preadsorbed ab98624) at 1:10000 and Goat anti-Mouse IgG H&L 680RD at 1:20000.
Blocking buffer: 3% milk in TBS-0.1% Tween® 20 (TBS-T).
Exposure time: 30 Sec.
All lanes: Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Human IgG1 (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] - Human IgG1 (Chimeric) ab323000) at 1/500 dilution
Lane 1: Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 µg
Lane 2: Recombinant SARS-COV-2 NSP2 protein (His tagged) at 0.2 µg
Lanes 1 - 2: Western blot - Goat Anti-Human IgG Fc (HRP) preadsorbed (Goat Anti-Human IgG Fc (HRP) preadsorbed ab98624) at 1/10000 dilution
Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 135 kDa
Exposure time: 30s
False colour image of Western blot: Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Human IgG1 (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 [8B12-C2] – Human IgG1 (Chimeric) ab322272) staining at 1/500 dilution, shown in black; Mouse Monoclonal 6X His tag® (ab18184) loading control staining at 1/200 dilution, shown in red. In Western blot, Anti-SARS-CoV-2 Spike Glycoprotein S1 [8B12-C2] – Human IgG1 (Chimeric) ab322272 was shown to bind specifically to SARS-CoV and SARS-CoV-2 Spike Glycoproteins. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Human IgG Fc (HRP) preadsorbed (Goat Anti-Human IgG Fc (HRP) preadsorbed ab98624) at 1:10000 and Goat anti-Mouse IgG H&L 680RD at 1:20000.
All lanes: Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 [8B12-C2] – Human IgG1 (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 [8B12-C2] – Human IgG1 (Chimeric) ab322272) at 1/500 dilution
Lane 1: Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 µg
Lane 2: SARS-CoV-2 (2019-nCoV) Spike S1+S2 ECD-His Recombinant Protein at 0.2 µg
Lane 3: SARS-CoV Spike S1+S2 ECD-His Recombinant Protein at 0.2 µg
Lanes 1 - 3: Western blot - Goat Anti-Human IgG Fc (HRP) preadsorbed (Goat Anti-Human IgG Fc (HRP) preadsorbed ab98624) at 1/10000 dilution
Lanes 1 - 3: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 120-160 kDa
Exposure time: 5s
Lane 1: Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active, His tagged) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 ug
Lane 2: SARS-CoV Spike S1+S2 ECD-His Recombinant Protein at 0.2 ug
Performed under reducing conditions.
Observed band size: 120-160 kDa.
False colour image of Western blot: Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) ab322999) staining at 1/500 dilution, shown in black; Mouse Monoclonal 6X His tag® (ab18184) loading control staining at 1/1000 dilution, shown in red. In Western blot, Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) ab322999 was shown to bind specifically to SARS-CoV and SARS-CoV-2 Spike Glycoproteins. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Chicken IgY H&L (HRP) preadsorbed (Goat Anti-Chicken IgY H&L (HRP) preadsorbed ab7118) at 1:10000 and Goat anti-Mouse IgG H&L 680RD at 1:20000.
Blocking buffer: 3% milk in TBS-0.1% Tween® 20 (TBS-T).
Exposure time: 2 min.
All lanes: Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) ab322999) at 1/500 dilution
Lane 1: Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) ab273068) at 0.2 µg
Lane 2: SARS-CoV Spike S1+S2 ECD-His Recombinant Protein at 0.2 µg
Lanes 1 - 2: Western blot - Goat Anti-Chicken IgY H&L (HRP) preadsorbed (Goat Anti-Chicken IgY H&L (HRP) preadsorbed ab7118) at 1/10000 dilution
Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 120-160 kDa
Exposure time: 2min
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