Product nameAnti-6X His tag® antibody - ChIP Grade
See all 6X His tag® primary antibodies
DescriptionRabbit polyclonal to 6X His tag® - ChIP Grade
SpecificityELISA: 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 applicationsSuitable for: IP, WB, IHC-P, ICC/IF, ELISA, ChIP, Flow Cytmore details
Species reactivityReacts with: Species independent
Synthetic peptide: HHHHHH conjugated to KLH.
- Purchase matching WB positive control:ENO1 full length protein
- ICC/IF: His tag expressed in HEK293 cells. WB: HeLa whole cell lysate transfected with His-tag. ChIP: HEK-293T cell line.
General notesThis antibody recognizes both C-terminal and N-terminal tags. HIS-TAG® is a trademark of EMD Biosciences, Inc.
See other anti-rabbit secondary antibodies that can be used with this antibody.
Storage instructionsShipped 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 bufferPreservative: 0.1% Sodium azide
Concentration information loading...
PurityImmunogen affinity purified
- Anti-6X His tag® antibody (ab14923)
- Anti-6X His tag® antibody [4D11] (HRP) (ab202004)
- Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
- Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
- Anti-6X His tag® antibody [JPAR-2] (ab245114)
- Anti-6X His tag® antibody [JPAR-2] - BSA and Azide free (ab245203)
- Anti-6X His tag® antibody [4D11] (ab5000)
ChIP Related Products
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.
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. PubMed: 21483675|
|ICC/IF||1/100 - 1/500.|
|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.
RelevanceThe 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 localizationDepends upon the localization of the parent protein tagged with hexahistidine.
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
ab9108 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).
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).
All lanes : Anti-6X His tag® antibody - ChIP Grade (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.
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
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
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) »
- Gomes JR et al. Delivery of an anti-transthyretin Nanobody to the brain through intranasal administration reveals transthyretin expression and secretion by motor neurons. J Neurochem 145:393-408 (2018). Read more (PubMed: 29527688) »