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...
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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.
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|
Dot blot |
Use at an assay dependent concentration.
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|
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 |
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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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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 -
ab5000 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).
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All 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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ab5000 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