Anti-DNA polymerase alpha antibody (ab31777)

Overview

  • Product name
    Anti-DNA polymerase alpha antibody
    See all DNA polymerase alpha primary antibodies
  • Description
    Rabbit polyclonal to DNA polymerase alpha
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Chicken, Human, Xenopus laevis
    Predicted to work with: Mouse, Rat, Dog
  • Immunogen

    Synthetic peptide derived from within residues 1100 - 1200 of Human DNA polymerase alpha.

    (Peptide available as ab30406.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: (WB) HeLa; Jurkat; A431: (IF) MCF7; HeLa; HEK293; HepG2

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped 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 buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 166 kDa).
IHC-P 1/100. Perform 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • DKFZp686K1672 antibody
    • DNA polymerase alpha 1 catalytic subunit antibody
    • DNA polymerase alpha catalytic subunit antibody
    • DNA polymerase alpha catalytic subunit p180 antibody
    • DNA polymerase alpha p180 subunit antibody
    • DPOLA_HUMAN antibody
    • OTTHUMP00000023089 antibody
    • OTTHUMP00000023090 antibody
    • P180 antibody
    • p180 subunit antibody
    • POLA antibody
    • POLA1 antibody
    • Polymerase (DNA directed), alpha 1, catalytic subunit antibody
    • Polymerase (DNA-directed), alpha (70kD) antibody
    see all

Images

  • 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 whole cell lysate (ab7899)
    Lane 3 : A431 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?)
  • 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

References

This product has been referenced in:
  • Yun J  et al. Reduced cohesin destabilizes high-level gene amplification by disrupting pre-replication complex bindings in human cancers with chromosomal instability. Nucleic Acids Res 44:558-72 (2016). Read more (PubMed: 26420833) »
  • Hashimoto Y  et al. Rad51 protects nascent DNA from Mre11-dependent degradation and promotes continuous DNA synthesis. Nat Struct Mol Biol 17:1305-11 (2010). Read more (PubMed: 20935632) »

See all 3 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Xenopus laevis Cell lysate - whole cell (Egg extract and replicating chromatin)
Specification
Egg extract and replicating chromatin
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Feb 17 2014

Thank you for your patience.
For ab57296, the sequence homology of the immunogen with mouse is 83% - which is good.
The lab let me know that for ab65009, the amino acid range of the immunogen is within residues 100-300 of human DNA Polymerase ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Chicken Cell lysate - nuclear (DT40)
Loading amount
50 µg
Specification
DT40
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Aug 14 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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