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
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) »
- 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) »