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ChIP - 6X His tag antibody - ChIP Grade (ab9108)

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Western blot - 6X His tag® antibody - ChIP Grade (ab9108)

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Western blot - 6X His tag antibody - ChIP Grade (ab9108)

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Product Name
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6X His tag® antibody - ChIP Grade
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See all 6X His tag antibodies (33)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to 6X His tag® - ChIP Grade
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Immunogen
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Synthetic peptide: HHHHHH conjugated to KLH. BLAST 'HHHHHH' with or 
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Specificity
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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.
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Tested applications
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ChIP, ELISA, Flow Cyt, ICC/IF, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions ChIP: ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. ELISA: 1/1000 (see PMID 19008226). Flow Cyt: Use at an assay dependent concentration (see Abreview published on 22nd January, 2007). ICC/IF: Use at a dilution of 1/100-1/500 for overnight incubation. WB: Western blotting up to 1/1000. IP: Use at an assay dependent concentration.
Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Depends upon the localization of the parent protein tagged with hexahistidine.
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies Tags & Cell Markers >> Epitope Tags >> His Tag
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Relevance
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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.
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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PBS with 0.1% sodium azide Material safety datasheet (MSDS) for this product: Sodium Azide MSDS
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Notes
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This antibody recognizes both C-terminal and N-terminal tags. HIS-TAG® is a trademark of EMD Biosciences, Inc.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this 6X His tag® antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for 6X His tag® antibody - ChIP Grade images, references, products related to ab9108 and other tools.
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6X His tag® antibody - ChIP Grade images:
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ChIP - 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).
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Western blot - 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
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Western blot - 6X His tag antibody - ChIP Grade (ab9108)
All lanes : 6X His tag® antibody - ChIP Grade (ab9108) at 1/1000 dilution
Lane 1 : 10ug non-transformed lysate Lane 2 : 10ug non-transformed lysate + 1mM IPTG Lane 3 : 10ug E.coli Lysate harbouring His-tag-RelA NTD, non-induced Lane 4 : 10ug Induction of His-tag RelA NTD 1 mM IPTG Lane 5 : 10ug Induction of His-tag RelA NTD 1 mM IPTG
Secondary HRP conjugated goat anti-rabbit IgG developed using the ECL technique
Performed under reducing conditions.
Observed band size : 37 kDa (why is the actual band size different from the predicted?) Additional bands at : 12 kDa (possible non-specific binding).
Exposure time : 1 minute
This image is courtesy of an Abreview.
See Abreview
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References for 6X His tag® antibody - ChIP Grade (ab9108)
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This product has been used in: (two most recent references)
Kalamajski S & Oldberg A Homologous Sequence in Lumican and Fibromodulin Leucine-rich Repeat 5-7 Competes for Collagen Binding. J Biol Chem 284:534-9 (2009). I-ELISA. PubMed: 19008226
Lei H et al. Caudal-like PAL-1 directly activates the bodywall muscle module regulator hlh-1 in C. elegans to initiate the embryonic muscle gene regulatory network. Development 136:1241-9 (2009). ChIP. PubMed: 19261701
See all 13 publication references for this product.
If you publish research using ab9108 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to 6X His tag®
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6X His tag® antibody - ChIP Grade - more information
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6X His tag® antibody - ChIP Grade - related products:
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Contact Abcam with a Technical Enquiry about 6X His tag® antibody - ChIP Grade |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"