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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


Images

Western blot - Anti-6X His tag® antibody [HIS.H8] (AB18184), expandable thumbnail
  • Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (AB18184), expandable thumbnail
  • Western blot - Anti-6X His tag® antibody [HIS.H8] (AB18184), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (AB18184), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (AB18184), expandable thumbnail

Publications

Key facts

Isotype
IgG2b
Host species
Mouse
Storage buffer

pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPELISADotWBICC/IF
Tag
Tested
Expected
Expected
Tested
Tested

Tested
Tested

Species
Tag
Dilution info
1/200
Notes

-

Expected
Expected

Species
Tag
Dilution info
Use at an assay dependent concentration.
Notes

-

Expected
Expected

Species
Tag
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Tag
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Tag
Dilution info
1/200
Notes

-

Associated Products

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Alternative names

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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

Key facts

Isotype
IgG2b
Form
Liquid
Clonality
Monoclonal
Clone number
HIS.H8
Purification technique
Affinity purification Protein A
Specificity

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.

Concentration
Loading...
Purification notes

Purified from TCS by Protein A affinity chromatography

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Rachel Loh

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    All lanes: IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time: 5min

  • Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Mr Chris Tracy

    Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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)

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Vladimir Milenkovic

    Western blot - 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

    Secondary

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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)

    Secondary

    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    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

    Secondary

    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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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