XRCC3 anticorps (X-Ray Repair Complementing Defective Repair in Chinese Hamster Cells 3)

Details for Product anti-XRCC3 Antibody No. ABIN151307
Antigène
  • CMM6
  • 4432412E01Rik
  • AI182522
  • AW537713
  • MGC69118
  • im:7142103
  • zgc:101608
  • si:dkey-11b8.1
  • xrcc3
  • XRCC3
  • X-ray repair cross complementing 3
  • X-ray repair complementing defective repair in Chinese hamster cells 3
  • X-ray repair complementing defective repair in Chinese hamster cells 3 L homeolog
  • XRCC3
  • Xrcc3
  • xrcc3.L
  • xrcc3
Reactivité
Humain
107
26
13
3
3
3
2
1
1
1
Hôte
Souris
91
20
3
Clonalité (Clone)
Monoclonal ()
Conjugué
Cet anticorp XRCC3 est non-conjugé
3
3
2
1
1
1
1
1
1
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (IHC), Western Blotting (WB)
87
40
33
19
11
6
4
3
3
2
1
1
Options
Immunogène His-tagged human XRCC3, over-expressed in E. coli [UniProt# O43542]
Clone 10F1-6
Isotype IgG1 kappa
Specificité This is specific for XRCC3. NB100-180 does not cross-react with Rad51B, Rad51C, Rad51D, Rad51, or XRCC2 in Western analysis.
Aucune reactivité croisée Souris
Purification Protein A purified
Plasmids, Primers & others Plasmids, Primers & others XRCC3 products on genomics-online (e.g. as negative or positive controls)
Antigène
Autre désignation XRCC3 (XRCC3 Antibody Extrait)
Sujet Gene Symbol: XRCC3
Poids moléculaire Theoretical MW: 38 kDa
ID gène 7517
UniProt O43542
Pathways Réparation de l'ADN
Indications d'application Western Blot 1:1000, Immunohistochemistry 1:10-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry-Paraffin 1:10-1:500This XRCC3 (10F1/6) antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections and Western blot. In WB, a band can be seen at ~38 kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Commentaires

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocole Protocol specific for XRCC3 Antibody Western Blot Protocol
1. Preparation of samples for loading Heat ~50-80ug of sample containing laemmli loading dye (containing SDS) at 90 °C for ~2 minutes.
. Load sample onto a 10 % Tris-HCL gel (Bio-Rad pre-cast) and run for ~30 minutes at 200V (or until dye front reaches bottom of gel).
. Place gel in transfer buffer for 10 minutes (192mM Glycine, 25mM Tris-HCL, 20 % Methanol). Pre-soak two pieces of Whatman paper and PVDF, as well. NOTE: The PVDF should be soaked in CH3OH for ~ 1minute, rinsed in ddH20 and then placed in transfer buffer.
. Transfer the protein from the gel to the membrane using a semi-dry transfer apparatus. Run for 20 minutes at 20V.
. Block non-specific proteins with blocking buffer #1 (10mM Tris-HCL pH 8.0, 300mM NaCL, 0.025 % Tween 20)for 10 minutes. Then continue blocking in blocking buffer #2 (buffer #1 + 15 % nonfat dry milk) for an additional hour, gently rocking at room temperature (RT) or overnight at 4 degrees Celcius.
. Dilute the primary antibody (anti-XRCC3, NB 100-180) in antibody dilution buffer (blocking buffer #1 + 2 % milk).
. Wash the membrane briefly with some blocking buffer #1 and then add your diluted primary antibody.
. Incubate the primary for 1 hour at room temperature, gently rocking. Again this can be done overnight at 4 Celsius.
. Wash 3X with blocking buffer #1 for 10 minutes, each, gently rocking.
. Incubate the diluted secondary antibody (anti-mouse IgG conjugated to HRP), diluted in antibody dilution buffer, for 1 hour at room temperature, gently rocking.
. Wash 2X with blocking buffer #1 for 10 minutes, each, gently rocking. Wash 1X with blocking buffer #1 for 30 minutes, gently rocking.
. Develop membrane with your chemiluminescent substrate. NOTE: HEK 293 and HeLa whole cell extracts have been used as positive controls for this antibody. Immunofluorescence Protocol
. Cells are grown on coverslips in a 6 well dish.
. Media is aspired off and cells are washed once with PBS.
. Cells are immersed in 100 % methanol for 5 minutes at -20 degrees Celcius.
. Cells are then blocked in PBS containing 4 % BSA overnight at 4 degrees Celcius.
. Cells are then washed 5 times for 5 minutes each with PBS.
. Cells are incubated with a 1:500 dilution of purified anti-XRCC3 [NB 100-180] for 1 hour at 37 degrees Celcius in a humid environment using a slide warmer.
. Cells are then washed 5 times for 5 minutes each with PBS.
. Cells are incubated with a diluted anti-mouse IgG Alexa 488 conjugated secondary antibody for 1 hour at 37 degrees Celcius in a humid environment using a slide warmer.
. Cells are then washed 5 times for 5 minutes each with PBS.
. Coverslips are mounted using Vectashield with DAPI and sealed with polyurethane and stored in the dark at 4 degrees Celcius.
. Visualization of immunostains was performed by confocal microscopy using a Leica TCS SP2 AOBS instrument.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer PBS ( pH 7.4)
Buffer contains: 0.1 % Sodium Azide
Agent conservateur Sodium azide
Précaution d'utilisation This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation Do not freeze.
Stock 4 °C
Stockage commentaire Store at 4°C. Do not freeze.
Images (Fournisseur)
Image no. 1 for anti-X-Ray Repair Complementing Defective Repair in Chinese Hamster Cells 3 (XRCC3) antibody (ABIN151307) Western Blot: XRCC3 Antibody (10F1/6) [ABIN151307SS] - XRCC3 detected in HEK293 lysat...
Produit citée dans: Chun, Buechelmaier, Powell: "Rad51 paralog complexes BCDX2 and CX3 act at different stages in the BRCA1-BRCA2-dependent homologous recombination pathway." dans: Molecular and cellular biology, Vol. 33, Issue 2, pp. 387-95, 2012 (PubMed). (Échantillon (espèces): Human).

Sage, Gildemeister, Knight: "Discovery of a novel function for human Rad51: maintenance of the mitochondrial genome." dans: The Journal of biological chemistry, Vol. 285, Issue 25, pp. 18984-90, 2010 (PubMed).

Shammas, Shmookler Reis, Koley, Batchu, Li, Munshi: "Dysfunctional homologous recombination mediates genomic instability and progression in myeloma." dans: Blood, Vol. 113, Issue 10, pp. 2290-7, 2009 (PubMed).

Gildemeister, Sage, Knight: "Cellular redistribution of Rad51 in response to DNA damage: novel role for Rad51C." dans: The Journal of biological chemistry, Vol. 284, Issue 46, pp. 31945-52, 2009 (PubMed).

Chan, Koritzinsky, Zhao, Bindra, Glazer, Powell, Belmaaza, Wouters, Bristow: "Chronic hypoxia decreases synthesis of homologous recombination proteins to offset chemoresistance and radioresistance." dans: Cancer research, Vol. 68, Issue 2, pp. 605-14, 2008 (PubMed).

Li, Jog, Reddy, Comai: "WRN controls formation of extrachromosomal telomeric circles and is required for TRF2DeltaB-mediated telomere shortening." dans: Molecular and cellular biology, Vol. 28, Issue 6, pp. 1892-904, 2008 (PubMed).

Compton, Choi, Cesare, Ozgür, Griffith: "Xrcc3 and Nbs1 are required for the production of extrachromosomal telomeric circles in human alternative lengthening of telomere cells." dans: Cancer research, Vol. 67, Issue 4, pp. 1513-9, 2007 (PubMed).

Bennett, Knight: "Cellular localization of human Rad51C and regulation of ubiquitin-mediated proteolysis of Rad51." dans: Journal of cellular biochemistry, Vol. 96, Issue 6, pp. 1095-109, 2005 (PubMed). (Échantillon (espèces): Human). Détails: Immunocytochemistry,Western Blotting,Immunofluorescence

Forget, Bennett, Knight: "Xrcc3 is recruited to DNA double strand breaks early and independent of Rad51." dans: Journal of cellular biochemistry, Vol. 93, Issue 3, pp. 429-36, 2004 (PubMed). (Échantillon (espèces): Human). Détails: Immunocytochemistry,Immunofluorescence

Fan, Kumaravel, Jalali, Marrano, Squire, Bristow: "Defective DNA strand break repair after DNA damage in prostate cancer cells: implications for genetic instability and prostate cancer progression." dans: Cancer research, Vol. 64, Issue 23, pp. 8526-33, 2004 (PubMed). (Échantillon (espèces): Human). Détails: Western Blotting

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