Overview

  • Product name
  • Description
    Rabbit polyclonal to XRCC1
  • Host species
    Rabbit
  • Specificity
    This antibody recognises human XRCC1 (x-ray cross complementing factor -1), a base excision repair protein.
  • Tested applications
    Suitable for: IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Horse, Guinea pig, Hamster, Cow, Dog, Pig, Chimpanzee, Ferret, Rhesus monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide (Human) conjugated to KLH - which represented aportion within the last 100 amino acids of the human X-ray Repair Cross Complementing Protein 1 (GenBank PID 11425942).

  • General notes
    Recently XRCC-1 has been shown to be physically associated with other DNA repair enzymes, including PARP, DNA ligase III and DNA polymerase beta.

Properties

Applications

Our Abpromise guarantee covers the use of ab9147 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
IP Use a concentration of 2 - 6 µg/ml.
WB 1/1000 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 70 kDa).
ICC/IF 1/200 - 1/500.

Target

  • Function
    Corrects defective DNA strand-break repair and sister chromatid exchange following treatment with ionizing radiation and alkylating agents.
  • Sequence similarities
    Contains 2 BRCT domains.
  • Post-translational
    modifications
    Phosphorylation of Ser-371 causes dimer dissociation. Phosphorylation by CK2 promotes interaction with APTX and APLF.
    Sumoylated.
  • Cellular localization
    Nucleus. Accumulates at sites of DNA damage.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA repair protein XRCC1 antibody
    • RCC antibody
    • X ray repair complementing defective repair in chinese hamster antibody
    • X ray Repair Complementing Defective Repair in Chinese Hamster Cells antibody
    • X ray repair complementing defective repair in chinese hamster cells 1 antibody
    • X ray repair cross complementing 1 antibody
    • X ray repair cross complementing protein 1 antibody
    • X ray repair, complementing defective, repair in Chinese hamster antibody
    • X-ray repair cross-complementing protein 1 antibody
    • XRCC 1 antibody
    • Xrcc1 antibody
    • XRCC1_HUMAN antibody
    see all

Images

  • All lanes : Anti-XRCC1 antibody (ab9147) at 1 µg/ml

    Lane 1 : Whole cell extract (50mcg) from mock treated HeLa cells.
    Lane 2 : Whole cell extract (50mcg) from siRNA treated HeLa cells.

    Predicted band size: 70 kDa
    Observed band size: 76,90 kDa
    why is the actual band size different from the predicted?



    Detection: ECL.
  • ab9147 staining human HeLa cells by ICC/IF.  Cells were PFA fixed and permeabilized in 0.5% Triton X100 prior to blocking in 5% BSA for 30 minutes at 20°C.  The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 20°C.  A Cy3® conjugated donkey anti-rabbit antibody diluted 1/400 was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Sun C  et al. Mobile phone signal exposure triggers a hormesis-like effect in Atm(+/+) and Atm(-/-) mouse embryonic fibroblasts. Sci Rep 6:37423 (2016). WB ; Mouse . Read more (PubMed: 27857169) »
  • Yousif AS  et al. Differential regulation of S-region hypermutation and class-switch recombination by noncanonical functions of uracil DNA glycosylase. Proc Natl Acad Sci U S A 111:E1016-24 (2014). Read more (PubMed: 24591630) »
See all 11 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Zebrafish Cell lysate - whole cell (Whole embyro 24hpf)
Gel Running Conditions
Non-reduced Denaturing (4-20%)
Loading amount
50 µg
Specification
Whole embyro 24hpf
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Mr. Stephen Moore

Verified customer

Submitted Dec 04 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (HeLa cells)
Specification
HeLa cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Alexander Rapp

Verified customer

Submitted Apr 18 2008

Question

> Could we get a detailed protocol from you, please? Harvest Cells: wash cells with pbs, add RIPA Buffer, refrigerate for 10-15 minutes until cell appear lysed, homogenize sample with 201/2-gauge needle, spin down and take off supernatant, quantitate protein concentration with the lowry method typically get 5-7 mg/ml from a 35 mm plate gel: load 30 micrograms of protein, 100v until through stack, 250v in resolving gel, semi-dry transfer to pvdf blotting: block for 1 hour in pbst + 5% milk XRCC1 polyclonal 1:5000 (ab9147-50) in pbst + 5% milk for 1.5 hours wash with pbst for 5 min, repeat 5 X Santa Cruz anti rabbit IgG hrp conjugate (sc2030) 1:3000 for 1.5 hours wash with pbst for 5 min, repeat X 5 expose to substrate for 1 min, put on film for 5 min i've used this protocol for about two years now on both helas and 293's. Also, please include the previous lot numbers that you used (if > you still have the information) and the Abcam order number or > purchase order number that was used for your latest order for > ab9147. unfortunately i don't have the previous lot numbers as i only use about 1 vial per year (i've found that i can re-use the primary 2-3 time with good results). The order reference number for the most recent lot is 19207 and the abcam order number is 53805. > > > > 1. Please describe the problem (high background, wrong band size, > more bands, no band etc). No signal at all, no background, nothing > 2. On what material are you testing the antibody in WB? cell extract from 293's > 3. How much protein did you load? 30 micrograms > â?¢How did you prepare the lysate for the analysis (protease > inhibitors etc)? wash cells with pbs, ripa buffer with PMSF, refrig. 10-15 min, homogenize with 201/2 gauge needle, spin down, take supernatant, quantitate protein > â?¢Did you heat the samples? yes > 4. Primary Antibody > â?¢Specification (in which species was it raised against)? human > â?¢At what dilution(s) have you tested this antibody? i normally use at 1:5000 > â?¢Incubation time, wash step? 1.5 hours, yes i wash 5 times, for 5 minn with pbst > > 5. Secondary Antibody > â?¢Specification (in which species was it raised against)? Santa Crus Anti-rabbit IgG-hrp SC2030 > â?¢At what dilution(s) have you tested this antibody? 1:3000 > â?¢Incubation time, wash step? 1.5 hr, yes, as above > â?¢Do you know whether the problems you are experiencing come > from the secondary? i've recently used the secondary with other polyclonal primaries with no problems (less than a week) > > 6. What detection method are you using? >ECL Western Blotting Analysis Kit from Amersham RPN2108 > 7. Background bands >NA â?¢Please provide an image of your blot (as an e-mail attachment, > a faxed image is not sufficient) see attached. > > 8. Optimization attempts > â?¢How many times have you tried the Western? I've been using this antibody successfully for two years, the most recent blots with the new antibody are my only failures. > > > 9. Did you apply positive and negative controls along with the > samples? Please specify. yes, i have positive controls on every blot (both nuclear extract from hela cells and whole cell extract from control 293 cells) Thanks

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Answer

Thank you for the details in which you have provided. Since you got absolutely no signal even with the HeLa nuclear extract (what is recommended as a positive control) it's definitely telling me that something is wrong with the batch/vial that you received. My suggestion would be to try increasing the concentration of the primary antibody, but since you're not even seeing any background I really don't think that would help. Anyway, I'm sending a replacement vial to you on order# 54735 and you should receive it tomorrow. Please let me know how it works out for you and if you have any further questions.

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