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Western blot
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Immunocytochemistry/ Immunofluorescence - Geminin antibody (ab12147)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Geminin antibody (ab12147)
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Product Name
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Geminin antibody
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See all Geminin antibodies (3)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Geminin
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human Geminin.
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Reacts with
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Hu
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Does not react with
(species key)
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Ms
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Specificity
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This antibody detects a band representing Geminin by Western blot at 33kD on 293T cell lysate, this band can be blocked by the immunising peptide. It also detects a band at about 45kD that is not blocked by the immunising peptide and is thought to be non-specific.
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Tested applications
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ICC/IF, IHC-P, WB
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Abreviews
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Application notes
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Recommended dilutions
ICC/IF: 1/400.
IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB: 1/500. Detects a band of approximately 33 kDa (predicted molecular weight: 23.5 kDa).
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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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> Other
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Relevance
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Geminin inhibits DNA replication by preventing the incorporation of MCM complex into prereplication complex (pre-RC). It is degraded during the mitotic phase of the cell cycle. It has a destruction box sequence (RRTLKVIQP) and is ubiquitinated by the anaphase promoting complex in vitro. Its destruction at the metaphase-anaphase transition permits replication in the succeeding cell cycle. Geminin interacts tightly with CDT1, a replication initiation factor necessary for MCM loading.
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Database links
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The links below go to external sites and will open in a new browser window |
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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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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 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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Batch dependent within range: 0.10 - 0.50 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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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Geminin antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Geminin antibody images, references, products related to ab12147 and other tools.
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Geminin antibody images:
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 Western blot
All lanes : Geminin antibody (ab12147) at 1/500 dilution
Lane 1 : 293 lysate
Lane 2 : 293 lysate with Geminin peptide (ab12408) at 1 µg/ml
Lysates/proteins at 5 µg/ml per lane.
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23.5 kDa
Observed band size : 33 kDa (why is the actual band size different from the predicted?)
The lower band represents Geminin (33 kDa - predicted molecular weight 23 kDa), and is blocked by the immunising peptide, the upper band is likely to be non-specific as it is not blocked by the peptide.
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Immunocytochemistry/ Immunofluorescence - Geminin antibody (ab12147)
ab12147 (1/400) staining geminin in assynchronous HeLa cells. Cells were fixed in paraformaldehyde and counterstained with DAPI in order to highlight the nucleus. Secondary antibody: goat anti-rabbit IgG conjugated to Cy3. Please refer to abreview for further details. This image is courtesy of an Abreview submitted by Dr Kirk McManus
See Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Geminin antibody (ab12147)
Image courtesy of Human Protein Atlas
ab12147 staining in human testis, showing staining of the cells in the ductus seminiferus (in brown). Paraffin embedded testis tissue was incubated with ab12147 (1:175 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab12147 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org .
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References for Geminin antibody (ab12147)
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This product has been used in:
Hochegger H et al. An essential role for Cdk1 in S phase control is revealed via chemical genetics in vertebrate cells. J Cell Biol 178:257-68 (2007). WB; Human. PubMed: 17635936
If you publish research using ab12147 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Geminin
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Geminin antibody - more information
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Customer reviews (feedback) regarding Geminin antibody |
Customer FAQs regarding Geminin antibody |
Protocols for Geminin antibody |
Price and availability of products related to Geminin antibody |
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Geminin antibody - related products:
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Geminin antibody - other tools:
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Contact Abcam with a Technical Enquiry about Geminin antibody |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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