Overview

  • Product name

    Anti-DNA polymerase alpha/POLA antibody
    See all DNA polymerase alpha/POLA primary antibodies
  • Description

    Rabbit polyclonal to DNA polymerase alpha/POLA
  • Host species

    Rabbit
  • 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 corresponding to Human DNA polymerase alpha/POLA aa 1100-1200.
    (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
  • General notes

     This product was previously labelled as DNA polymerase alpha

     

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/POLA 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
    All lanes : 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/POLA 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:

  • Abdel-Samad R  et al. Mechanism of action of the atypical retinoid ST1926 in colorectal cancer: DNA damage and DNA polymerase a. Am J Cancer Res 8:39-55 (2018). Read more (PubMed: 29416919) »
  • Bellelli R  et al. Pole Instability Drives Replication Stress, Abnormal Development, and Tumorigenesis. Mol Cell 70:707-721.e7 (2018). Read more (PubMed: 29754823) »
See all 7 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Feb 17 2014

Answer

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 alpha with UniProt accession number P09884. The alignment of the immunogen sequence with the mouse protein (P33609) gives only a 61.5% sequence identity.
We usually say that a homology of 85% or higher is best for cross-reactivity. Although the immunogen region is 50 residues, the actual epitope of the antibody binding site is only 5-10 residues. This could mean the actual homology for the epitope could be either better than 61.5% or could also be worse. This is very difficult to predict and would need to be tried out.
Since none of our Anti-DNA polymerase alpha antibody has been tested in mouse, you would be eligible for our testing discount program.
For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the to-be-tested product at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount in the amount of the price you paid for the to-be-tested product. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.
Please let me know if you would like to try any of the antibody and if I should send you a discount code.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Western blot
Sample
Chicken Cell lysate - nuclear (DT40)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
50 µg
Specification
DT40
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Aug 14 2009

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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