Anti-6X His tag® antibody [HIS.H8] (ab18184)
![Anti-6X His tag® antibody [HIS.H8] (ab18184)](https://www.abcam.com/ps/products/18/ab18184/Images/ab18184-26832-anti-6x-his-tag-antibody-hish8-immunoprecipitation.jpg "Anti-6X His tag® antibody [HIS.H8] (ab18184)")
Key features and details
- Mouse monoclonal [HIS.H8] to 6X His tag®
- Suitable for: ICC/IF, ELISA, Dot blot, WB, IP
- Reacts with: Species independent
- Isotype: IgG2b
Overview
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Product name
Anti-6X His tag® antibody [HIS.H8]
See all 6X His tag® primary antibodies -
Description
Mouse monoclonal [HIS.H8] to 6X His tag® -
Host species
Mouse -
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.
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Tested applications
Suitable for: ICC/IF, ELISA, Dot blot, WB, IPmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
6x His synthetic peptide.
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Positive control
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General notes
This product was changed from ascites to tissue culture supernatant on 5th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
HIS-TAG® is a trademark of EMD Biosciences, Inc.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Purification notes
Purified from TCS by Protein A affinity chromatography -
Clonality
Monoclonal -
Clone number
HIS.H8 -
Isotype
IgG2b -
Research areas
Images
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Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184) This image is courtesy of an Abreview submitted by Mr Chris Tracy
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. -
![Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)](https://www.abcam.com/ps/products/18/ab18184/Images/ab18184-261409-anti-6x-his-tag-antibody-hish8-western-blot.jpg)
Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184) This image is courtesy of an Abreview submitted by Vladimir MilenkovicAll lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution
Lane 1 : ARPE-19 whole cell lysate - non-transfected
Lane 2 : ARPE-19 whole cell lysate - transfected with rRab27a-His
Lysates/proteins at 20 µg per lane.
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: 5 minutesBlocked 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)](https://www.abcam.com/ps/products/18/ab18184/Images/ab18184-290288-anti-6x-his-tag-antibody-hish8-immunofluorescence.jpg)
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).
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![Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)](https://www.abcam.com/ps/products/18/ab18184/Images/ab18184-261408-anti-6x-his-tag-antibody-hish8-western-blot.jpg)
Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184) This image is courtesy of an Abreview submitted by Rachel LohAll lanes : 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: 5 minutes
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![Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)](https://www.abcam.com/ps/products/18/ab18184/Images/ab18184-26833-anti-6x-his-tag-antibody-hish8-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184) This image is courtesy of an anonymous abreview.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).
