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Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> DNA Polymerases
Overview
Product name
Anti-DNA polymerase alpha antibody
DNA polymerase alpha 抗体 (3件) 一覧
Description
Rabbit polyclonal to DNA polymerase alpha
Tested applications
WB, IHC-P, ICC/IFmore details
Cross reactivity
Reacts with
Chicken, Human
Predicted to work with
Mouse, Rat, Dog, Xenopus laevis
Immunogen
Synthetic peptide derived from within residues 1100 - 1200 of Human DNA polymerase alpha.
(Peptide available as ab30406.)
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
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> DNA Polymerases
Applications
Show applications key
Our Abpromise guarantee covers the use of ab31777 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 - 5 µg/ml.Detects a band of approximately 180 kDa (predicted molecular weight: 166 kDa).
IHC-P: 1/100Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use a concentration of 1 µg/ml
Target
Function
Plays an essential role in the initiation of DNA replication. During the S phase of the cell cycle, the DNA polymerase alpha complex (composed of a catalytic subunit POLA1/p180, a regulatory subunit POLA2/p70 and two primase subunits PRIM1/p49 and PRIM2/p58) is recruited to DNA at the replicative forks via direct interactions with MCM10 and WDHD1. The primase subunit of the polymerase alpha complex initiates DNA synthesis by oligomerising short RNA primers on both leading and lagging strands. These primers are initially extended by the polymerase alpha catalytic subunit and subsequently transferred to polymerase delta and polymerase epsilon for processive synthesis on the lagging and leading strand, respectively. The reason this transfer occurs is because the polymerase alpha has limited processivity and lacks intrinsic 3' exonuclease activity for proofreading error, and therefore is not well suited for replicating long complexes.
Sequence similarities
Belongs to the DNA polymerase type-B family.
Post-translational
modifications
A 165 kDa form is probably produced by proteolytic cleavage at Lys-124.
Cellular localization
Nucleus.
Target information above from: UniProt accessionP09884
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- DKFZp686K1672 antibody
- DNA polymerase alpha 1 catalytic subunit antibody
- DNA polymerase alpha catalytic subunit antibody
see all
Database links
- Entrez Gene: 418593 Chicken
- Entrez Gene: 5422 Human
- Entrez Gene: 18968 Mouse
- Entrez Gene: 85241 Rat
- Entrez Gene: 398200 Xenopus laevis
- Omim: 312040 Human
- SwissProt: P09884 Human
- SwissProt: P33609 Mouse
see all
Anti-DNA polymerase alpha antibody images:
Western blot - DNA polymerase alpha antibody (ab31777)
All lanes : Anti-DNA polymerase alpha antibody (ab31777) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 166 kDa
Observed band size : 180 kDa (why is the actual band size different from the predicted?)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DNA polymerase alpha antibody (ab31777)
Image courtesy of Human Protein Atlas ab31777 staining DNA polymerase alpha in human thyroid gland, showing a strong staining in nuclei of the glandular cells. Paraffin embedded human thyroid tissue was incubated with ab31777 (1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab31777 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
Immunocytochemistry/ Immunofluorescence - DNA polymerase alpha antibody (ab31777)
ICC/IF image of ab31777 stained human MCF7 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab31777, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and HepG2 cells.
References for Anti-DNA polymerase alpha antibody (ab31777)
This product has been referenced in:
- Ferguson RL & Maller JL Centrosomal localization of cyclin E-Cdk2 is required for initiation of DNA synthesis. Curr Biol 20:856-60 (2010). WB; Chinese Hamster.Read more (PubMed: 20399658) »
See 1 publication for this product
Publishing research using ab31777? 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"