Anti-6X His tag® antibody [4D11] (ab5000)
![Western blot - Anti-6X His tag® antibody [4D11] (ab5000)](https://www.abcam.com/ps/products/5/ab5000/Images/ab5000-9868-anti-6x-his-tag-antibody-4d11-western-blot.jpg "Western blot - Anti-6X His tag® antibody [4D11] (ab5000)")
Key features and details
- Mouse monoclonal [4D11] to 6X His tag®
- Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cyt
- Reacts with: Species independent
- Isotype: IgG2a
Related conjugates and formulations
Overview
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Product name
Anti-6X His tag® antibody [4D11]
See all 6X His tag® primary antibodies -
Description
Mouse monoclonal [4D11] to 6X His tag® -
Host species
Mouse -
Tested applications
Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cytmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Epitope
This antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50). -
General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 6.97% L-Arginine, PBS -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
4D11 -
Myeloma
NS1 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5000 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| AP |
Use at an assay dependent concentration.
|
|
| Dot blot |
Use at an assay dependent concentration.
|
|
| ELISA |
Use at an assay dependent concentration.
|
|
| WB | (2) |
1/1000.
|
| IP | (1) |
Use at an assay dependent concentration.
See Abreview. |
| ICC/IF | (1) |
Use at an assay dependent concentration.
|
| Flow Cyt |
Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
AP
Use at an assay dependent concentration. |
|
Dot blot
Use at an assay dependent concentration. |
|
ELISA
Use at an assay dependent concentration. |
|
WB
1/1000. |
|
IP
Use at an assay dependent concentration. See Abreview. |
|
ICC/IF
Use at an assay dependent concentration. |
|
Flow Cyt
Use at an assay dependent concentration. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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Relevance
The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. His-tags can be fused to the amino- or carboxy- termini of proteins in transfected or transformed cells. -
Cellular localization
Depends upon the localization of the parent protein tagged with hexahistidine. -
Alternative names
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
see all
Images
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Western blot - Anti-6X His tag® antibody [4D11] (ab5000)Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg
Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 30 seconds -
![Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)](https://www.abcam.com/ps/products/5/ab5000/Images/ab5000-9867-anti-6x-his-tag-antibody-4d11-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovicab5000 staining Human HEK 293 cells transfected with CACNB3-His by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% serum for 20 minutes at 25°C. The primary antibody (1/250 in 1% goat serum, 0.1% TX100, 1X PBS) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat anti-mouse antibody (1/500) was used as the secondary. DAPI was used for nuclei staining (blue). -
![Western blot - Anti-6X His tag® antibody [4D11] (ab5000)](https://www.abcam.com/ps/products/5/ab5000/Images/ab5000-9870-anti-6x-his-tag-antibody-4d11-western-blot.jpg)
Western blot - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir MilenkovicAll lanes : Anti-6X His tag® antibody [4D11] (ab5000) at 1/2000 dilution
Lane 1 : Lysates prepared from HEK-293 cells
Lane 2 : Lysates prepared from HEK-293 cells transfected with Beta3-His (CACNB3)
Secondary
All lanes : HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution
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![Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)](https://www.abcam.com/ps/products/5/ab5000/Images/ab5000-9869-anti-6x-his-tag-antibody-4d11-immunoprecipitation.jpg)
Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovicab5000 Immunoprecipitate in human HEK293 whole cell lysate. 25µg of cell lysate incubated with primary antibody (undiluted) and matrix (Protein G) for 16 hours at 4°C. For western blotting an undiluted HRP-Goat anti-rabbit IgG polyclonal was used. Beta3-His fusion protein was immunoprecipitated using anti His Ab, and specific beta3a band (56kDa) was detected using anti beta3 Ab
Lane 1. Lysate of HEK293 cells expressing beta3-His (CACNB3)
Lane 2. IP with anti His Ab
Lane 3. Non bound fraction
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (30)
ab5000 has been referenced in 30 publications.
- Peña-Oyarzun D et al. PKD2/polycystin-2 induces autophagy by forming a complex with BECN1. Autophagy 17:1714-1728 (2021). PubMed: 32543276
- Chen C et al. SUMOylation promotes extracellular vesicle-mediated transmission of lncRNA ELNAT1 and lymph node metastasis in bladder cancer. J Clin Invest 131:N/A (2021). PubMed: 33661764
- Rajan S et al. A simple, rapid, and sensitive fluorescence-based method to assess triacylglycerol hydrolase activity. J Lipid Res 62:100115 (2021). PubMed: 34508728
- Reichel C et al. Detection of black market follistatin 344. Drug Test Anal 12:1522-1533 (2020). PubMed: 33460286
- Lv Z et al. CD19-targeting fusion protein combined with PD1 antibody enhances anti-tumor immunity in mouse models. Oncoimmunology 9:1747688 (2020). PubMed: 32363119