Anti-6X His tag® antibody (ab9108)
- Datasheet
- References (88)
- Protocols
Overview
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Product name
Anti-6X His tag® antibody
See all 6X His tag® primary antibodies -
Description
Rabbit polyclonal to 6X His tag® -
Host species
Rabbit -
Specificity
ELISA: Antibody specificity was verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA). A 1:25,000 dilution of the antibody gave an OD=1.0 in a 15 minute reaction using HRP-conjugated Goat anti-Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run. -
Tested applications
Suitable for: IP, WB, IHC-P, ICC/IF, ELISA, ChIP, Flow Cytmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide: HHHHHH conjugated to KLH.
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Positive control
- Purchase matching WB positive control:Recombinant ENO1 protein
- ICC/IF: His tag expressed in HEK293 cells. WB: HeLa whole cell lysate transfected with His-tag. ChIP: HEK-293T cell line.
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General notes
This antibody recognizes both C-terminal and N-terminal tags. HIS-TAG® is a trademark of EMD Biosciences, Inc.Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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: 0.1% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab9108 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 |
|---|---|---|
| IP | Use at an assay dependent concentration. | |
| WB | 1/1000. | |
| IHC-P | Use at an assay dependent concentration. PubMed: 21483675 | |
| ICC/IF | 1/100 - 1/500. | |
| ELISA | 1/1000. | |
| ChIP | Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. | |
| Flow Cyt | Use at an assay dependent concentration. (see Abreview published on 22nd January, 2007). ab171870 - Rabbit polyclonal IgG, 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. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted 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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Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody - ChIP Grade (ab9108)This image is courtesy of Vladimir Milenkovicab9108 staining CACNB3 with His tag expressed in HEK293 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum, 0.1% TX100 and PBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/750).
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ChIP - Anti-6X His tag® antibody - ChIP Grade (ab9108)A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach). -
Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)This image is courtesy of an anonymous AbreviewAll lanes : Anti-6X His tag® antibody (ab9108) at 1/4000 dilution
Lane 1 : HeLa whole cell lysate - transfected with empty vector
Lane 2 : HeLa whole cell lysate - transfected with His-tagged gene
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
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Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)Use of ab9108 in western blotting at 1:1000 against Epitope Tag Positive Control lysate (ab4524).
Lane 1: 2 µl lysate
Lane 2: 1 µl lysate
Lane 3: 0.5 µl lysate
Lane 4: 0.25 µl lysate Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate Lane 3: 0.5 µl lysate Lane 4: 0.25 µl lysate
Protocols
References
This product has been referenced in:
- Diakonov EE et al. Evidences against vesicle-dependent trafficking and involvement of extracellular proteasomes into cell-to-cell communications. Biochem Biophys Res Commun 508:368-373 (2019). Read more (PubMed: 30503341) »
- Müskens FM et al. Design, Synthesis, and Evaluation of a Diazirine Photoaffinity Probe for Ligand-Based Receptor Capture Targeting G Protein-Coupled Receptors. Mol Pharmacol 95:196-209 (2019). Read more (PubMed: 30514721) »
