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Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> Other
Overview
Product name
Anti-Geminin antibody
See all Geminin products (3) ...
Description
Rabbit polyclonal to Geminin
Specificity
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.
Tested applications
ICC/IF, WB, IHC-Pmore details
Cross reactivity
Reacts with
Human
Does not react with
Mouse
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human Geminin.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> Other
Applications
Show applications key
Our Abpromise guarantee covers the use of ab12147 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: 1/400
WB: 1/500Detects a band of approximately 33 kDa (predicted molecular weight: 23.5 kDa).
IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Target
Function
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. Its destruction at the metaphase-anaphase transition permits replication in the succeeding cell cycle.
Sequence similarities
Belongs to the geminin family.
Developmental stage
Absent during G1 phase, accumulates during S, G2, and M phases, and disappears at the time of the metaphase-anaphase transition.
Target information above from: UniProt accessionO75496
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- DNA replication inhibitor antibody
- Gem antibody
- GEMI_HUMAN antibody
see all
Database links
- Entrez Gene: 51053 Human
- Omim: 602842 Human
- SwissProt: O75496 Human
- Unigene: 234896 Human
Anti-Geminin antibody images:
Western blot
All lanes : Anti-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.
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
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
.Western blot - Geminin antibody (ab12147)
Lane 1 : Anti-Geminin antibody (ab12147) at 1 µg/ml
Lane 2 : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1 µg/ml
Lane 1 : Geminin protein (His tag) (ab40570)
Lane 2 : Geminin protein (His tag) (ab40570)
Lysates/proteins at 0.1 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 23.5 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)
Western blot - Geminin antibody (ab12147)
Anti-Geminin antibody (ab12147) at 1/500 dilution + Cell lysate prepared from human HeLa nuclear extract at 50 µg
Secondary
HRP-conjugated goat poyclonal to rabbit IgG at 1/5000 dilution
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?)
Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 10 minutes
This image is a courtesy of Anonymous Abreview
References for Anti-Geminin antibody (ab12147)
This product has been referenced in:
- Hochegger Het 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.Read more (PubMed: 17635936) »
See 1 publication for this product
Publishing research using ab12147? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"