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AB65963

Anti-XPA antibody [5F12]

4

(1 Review)

|

(12 Publications)

Mouse Monoclonal XPA antibody. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 12 publications. Immunogen corresponding to Recombinant Full Length Protein corresponding to Human XPA.

View Alternative Names

XPAC, XPA, DNA repair protein complementing XP-A cells, Xeroderma pigmentosum group A-complementing protein

4 Images
Immunocytochemistry/ Immunofluorescence - Anti-XPA antibody [5F12] (AB65963)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-XPA antibody [5F12] (AB65963)

Immunofluorescent staining of human fibroblast cells (GM0637) using ab65963 at a dilution of 1/100. The cells were non-irradiated (left) or irradiated with UV at 20 J/m2 (right), fixed after 30 minutes with paraformaldehyde and counter-stained with Hoescht. The top panels show the antibody staining, the middle panels show the Hoescht staining and the bottom panels are a merge of the two images. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-mouse IgG, used at a dilution of 1/5000.

Western blot - Anti-XPA antibody [5F12] (AB65963)
  • WB

Lab

Western blot - Anti-XPA antibody [5F12] (AB65963)

Lanes 1-3 : Merged signal (red and green). Green - ab65963 observed at 38 kDa. Red - loading control, ab52866 observed at 50 kDa.

ab65963 Anti-XPA antibody [5F12] was shown to specifically react with XPA in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264663 (knockout cell lysate ab258764) was used. Wild-type and XPA knockout samples were subjected to SDS-PAGE. ab65963 and Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-XPA antibody [5F12] (ab65963) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

XPA knockout HeLa cell lysate at 20 µg

Lane 3:

293T cell lysate at 20 µg

Predicted band size: 31 kDa

Observed band size: 38 kDa

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Western blot - Anti-XPA antibody [5F12] (AB65963)
  • WB

Supplier Data

Western blot - Anti-XPA antibody [5F12] (AB65963)

All lanes:

Western blot - Anti-XPA antibody [5F12] (ab65963) at 1/2000 dilution

Lane 1:

HeLa cell extract (XPA wild type)

Lane 2:

XP12ROSV cell extract (XPA deficient)

Predicted band size: 31 kDa

false

Western blot - Anti-XPA antibody [5F12] (AB65963)
  • WB

Unknown

Western blot - Anti-XPA antibody [5F12] (AB65963)

All lanes:

Western blot - Anti-XPA antibody [5F12] (ab65963) at 0.1 µg/mL

All lanes:

Crude extract of HeLa cells at 200 µg

Predicted band size: 31 kDa

Observed band size: 38 kDa

false

Key facts

Host species

Mouse

Clonality

Monoclonal

Clone number

5F12

Isotype

IgG2b

Carrier free

No

Reacts with

Human

Applications

ICC/IF, WB

applications

Immunogen

Recombinant Full Length Protein corresponding to Human XPA.

P23025

Epitope

Amino acids 30-47.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "0.1-1 µg/mL", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Mouse": { "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }
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Properties and storage information

Form
Liquid
Purification technique
Ion exchange chromatography
Storage buffer
pH: 6 - 8.5 Constituents: PBS, 50% Glycerol (glycerin, glycerine)
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Storage information
Stable for 12 months at -20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The XPA (xeroderma pigmentosum group A) protein also known as DNA repair protein complementing XP-A cells is a vital player in the nucleotide excision repair (NER) machinery. It has a molecular mass of approximately 31 kDa. XPA functions as a DNA-binding protein and is expressed in various tissues including skin cells. XPA recognizes and stabilizes the DNA damage site acting as a scaffold to recruit other repair factors ensuring the proper assembly of essential components in the repair complex.
Biological function summary

The protein is essential for maintaining genomic stability by facilitating the repair of DNA lesions. XPA operates within a multiprotein complex that includes the proteins ERCC1 and XPF. The complex identifies a wide range of structural abnormalities in DNA such as UV-induced photoproducts and chemically induced adducts. Without XPA the NER system's efficiency decreases leaving cells vulnerable to genotoxic stresses causing mutations and damage accumulation.

Pathways

XPA plays a role in the nucleotide excision repair and the base excision repair pathways. These pathways work jointly to ensure the removal of damaged DNA sections and accurate DNA synthesis. Within the NER pathway XPA interacts with proteins like RPA (Replication Protein A) and TFIIH (Transcription Factor IIH) to mediate the excision of damaged oligonucleotides. Its activity is critical for preserving the integrity of the genetic code especially in response to DNA damage caused by external agents like UV radiation.

Mutations or deficiencies in XPA are linked to xeroderma pigmentosum a disorder characterized by extreme sensitivity to ultraviolet light and a heightened risk of skin cancer. This condition results from impaired DNA repair leading to the accumulation of unrepaired DNA damage. XPA also interacts with other NER proteins like ERCC1 and XPF whose dysfunction is implicated in similar DNA repair disorders emphasizing the interconnected nature of these repair pathways in protecting against genotoxic agents.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Involved in DNA nucleotide excision repair (NER). Initiates repair by binding to damaged sites with various affinities, depending on the photoproduct and the transcriptional state of the region. Required for UV-induced CHEK1 phosphorylation and the recruitment of CEP164 to cyclobutane pyrimidine dimmers (CPD), sites of DNA damage after UV irradiation (PubMed : 19197159). During NER stimulates the 5'-3' helicase activity of XPD/ERCC2 and the DNA translocase activity of XPB/ERCC3 (PubMed : 31253769). Connects XPD/ERCC2 and XPB/ERCC3 during NER, retaining DNA near the XPB/ERCC3 active site, and stabilizing the complex in a different conformation than in transcribing TFIIH (PubMed : 31253769).
See full target information XPA
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Publications (12)

Recent publications for all applications. Explore the full list and refine your search

Nucleic acids research 53: PubMed40744501

2025

Genome-wide mapping of formaldehyde-induced DNA-protein crosslinks reveals unique patterns of formation and transcription-coupled removal in mammalian cells.

Applications

Unspecified application

Species

Unspecified reactive species

Duha Alshareef,Charlie T Nguyen,Kayla N Tucker,Micah D Gearhart,Natalia Y Tretyakova,Colin Campbell

Molecular cell 84:1224-1242.e13 PubMed38458201

2024

CDK-independent role of D-type cyclins in regulating DNA mismatch repair.

Applications

Unspecified application

Species

Unspecified reactive species

Gergely Rona,Bearach Miwatani-Minter,Qingyue Zhang,Hailey V Goldberg,Marc A Kerzhnerman,Jesse B Howard,Daniele Simoneschi,Ethan Lane,John W Hobbs,Elizabeth Sassani,Andrew A Wang,Sarah Keegan,Daniel J Laverty,Cortt G Piett,Lorinc S Pongor,Miranda Li Xu,Joshua Andrade,Anish Thomas,Piotr Sicinski,Manor Askenazi,Beatrix Ueberheide,David Fenyö,Zachary D Nagel,Michele Pagano

Nucleic acids research 51:12174-12184 PubMed37843153

2023

Ubiquitin signaling and the proteasome drive human DNA-protein crosslink repair.

Applications

Unspecified application

Species

Unspecified reactive species

Maram Essawy,Lisa Chesner,Duha Alshareef,Shaofei Ji,Natalia Tretyakova,Colin Campbell

DNA repair 109:103260 PubMed34883264

2021

XPA is susceptible to proteolytic cleavage by cathepsin L during lysis of quiescent cells.

Applications

Unspecified application

Species

Unspecified reactive species

Saman Khan,William Cvammen,Nadeen Anabtawi,Jun-Hyuk Choi,Michael G Kemp

Nature communications 12:585 PubMed33500419

2021

The Bloom syndrome complex senses RPA-coated single-stranded DNA to restart stalled replication forks.

Applications

Unspecified application

Species

Unspecified reactive species

Ann-Marie K Shorrocks,Samuel E Jones,Kaima Tsukada,Carl A Morrow,Zoulikha Belblidia,Johanna Shen,Iolanda Vendrell,Roman Fischer,Benedikt M Kessler,Andrew N Blackford

Pathogens (Basel, Switzerland) 8: PubMed31779191

2019

β-HPV 8E6 Attenuates ATM and ATR Signaling in Response to UV Damage.

Applications

Unspecified application

Species

Unspecified reactive species

Jazmine A Snow,Vaibhav Murthy,Dalton Dacus,Changkun Hu,Nicholas A Wallace

DNA repair 52:70-80 PubMed28237621

2017

Inorganic arsenic inhibits the nucleotide excision repair pathway and reduces the expression of XPC.

Applications

Unspecified application

Species

Unspecified reactive species

Nathaniel Holcomb,Mamta Goswami,Sung Gu Han,Tim Scott,John D'Orazio,David K Orren,C Gary Gairola,Isabel Mellon

Oncotarget 7:38367-38379 PubMed27223263

2016

Targeting DNA repair with aphidicolin sensitizes primary chronic lymphocytic leukemia cells to purine analogs.

Applications

Unspecified application

Species

Unspecified reactive species

Eliza Starczewska,Maxime Beyaert,Lucienne Michaux,Marie-Christiane Vekemans,Pascale Saussoy,Vanesa Bol,Ainhoa Arana Echarri,Caroline Smal,Eric Van Den Neste,Françoise Bontemps

The Journal of biological chemistry 291:5309-19 PubMed26733197

2016

Base and Nucleotide Excision Repair of Oxidatively Generated Guanine Lesions in DNA.

Applications

Unspecified application

Species

Unspecified reactive species

Vladimir Shafirovich,Konstantin Kropachev,Thomas Anderson,Zhi Liu,Marina Kolbanovskiy,Brooke D Martin,Kent Sugden,Yoonjung Shim,Xuejing Chen,Jung-Hyun Min,Nicholas E Geacintov

PloS one 8:e55493 PubMed23405159

2013

An inverse switch in DNA base excision and strand break repair contributes to melphalan resistance in multiple myeloma cells.

Applications

WB

Species

Human

Mirta M L Sousa,Kamila Anna Zub,Per Arne Aas,Audun Hanssen-Bauer,Aida Demirovic,Antonio Sarno,Erming Tian,Nina B Liabakk,Geir Slupphaug
View all publications
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