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Read our guarantee »Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> Nucl. Excision Repair
Anti-XPC antibody [3.26]
See all XPC products (5) ...
Mouse monoclonal [3.26] to XPC
ICC/IF, IHC-P, Flow Cyt, WBmore details
Reacts with
Human
Recombinant full length human XPC protein expressed in E. coli.
Raji whole cell lysate, HeLa cell lysate.
Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: None
Constituents: PBS
Concentration information loading...
Protein G purified
Purified from tissue culture supernatant.
Monoclonal
3.26
NS1
IgG1
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> Nucl. Excision Repair
Our Abpromise guarantee covers the use of ab6264 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: 1/100
IHC-P: 1/100Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt: Use 1µl for 106 cells.
WB: 1/500 - 1/1000.Detects a band of approximately 113-120 kDa (predicted molecular weight: 105 kDa).(Block with 1% BSA instead of milk.)
Involved in global genome nucleotide excision repair (GG-NER) by acting as damage sensing and DNA-binding factor component of the XPC complex. Has only a low DNA repair activity by itself which is stimulated by RAD23B and RAD23A. Has a preference to bind DNA containing a short single-stranded segment but not to damaged oligonucleotides. This feature is proposed to be related to a dynamic sensor function: XPC can rapidly screen duplex DNA for non-hydrogen-bonded bases by forming a transient nucleoprotein intermediate complex which matures into a stable recognition complex through an intrinsic single-stranded DNA-binding activity.
The XPC complex is proposed to represent the first factor bound at the sites of DNA damage and together with other core recognition factors, XPA, RPA and the TFIIH complex, is part of the pre-incision (or initial recognition) complex. The XPC complex recognizes a wide spectrum of damaged DNA characterized by distortions of the DNA helix such as single-stranded loops, mismatched bubbles or single stranded overhangs. The orientation of XPC complex binding appears to be crucial for inducing a productive NER. XPC complex is proposed to recognize and to interact with unpaired bases on the undamaged DNA strand which is followed by recruitment of the TFIIH complex and subsequent scanning for lesions in the opposite strand in a 5'-to-3' direction by the NER machinery. Cyclobutane pyrimidine dimers (CPDs) which are formed upon UV-induced DNA damage esacpe detection by the XPC complex due to a low degree of structural perurbation. Instead they are detected by the UV-DDB complex which in turn recruits and cooperates with the XPC complex in the respective DNA repair. In vitro, the XPC:RAD23B dimer is sufficient to initiate NER; it preferentially binds to cisplatin and UV-damaged double-stranded DNA and also binds to a variety of chemically and structurally diverse DNA adducts. XPC:RAD23B contacts DNA both 5' and 3' of a cisplatin lesion with a preference for the 5' side. XPC:RAD23B induces a bend in DNA upon binding. XPC:RAD23B stimulates the activity of DNA glycosylases TDG and SMUG1.
Defects in XPC are a cause of xeroderma pigmentosum complementation group C (XP-C) [MIM:278720]; also known as xeroderma pigmentosum III (XP3). XP-C is a rare human autosomal recessive disease characterized by solar sensitivity, high predisposition for developing cancers on areas exposed to sunlight and, in some cases, neurological abnormalities.
Belongs to the XPC family.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Ubiquitinated upon UV irradiation; the ubiquitination requires the UV-DDB complex, appears to be reversible and does not serve as a signal for degradation.
Nucleus. Cytoplasm. Omnipresent in the nucleus and consistently associates with and dissociates from DNA in the absence of DNA damage. Continuously shuttles between the cytoplasm and the nucleus, which is impeded by the presence of NER lesions.
Target information above from: UniProt accessionQ01831
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Western blot - XPC antibody [3.26] (ab6264)
![Western blot - XPC antibody [3.26] (ab6264)](/ps/datasheet/Images/6/ab6264/ab6264_3.jpg)
Lane 1 : Anti-XPC antibody [3.26] (ab6264) at 1/1000 dilution
Lane 2 : Anti-XPC antibody [3.26] (ab6264) at 1/500 dilution
Lane 1 : Raji whole cell extract
Lane 2 : Raji whole cell extract
Predicted band size : 105 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
Immunocytochemistry/ Immunofluorescence - XPC antibody [3.26] (ab6264)
![Immunocytochemistry/ Immunofluorescence - XPC antibody [3.26] (ab6264)](/ps/datasheet/Images/6/ab6264/ab6264_4.jpg)
Ab6264 staining human XPC in HeLa cells (left), and DAPI staining (right) by immunofluorescence.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-XPC antibody [3.26](ab6264)
](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-1.jpg)
ab6264 (4µg/ml) staining XPC in human tonsil, using an automated system (DAKO Autostainer Plus). Using this protocol there is weak nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Flow Cytometry - XPC antibody [3.26] (ab6264)
![Flow Cytometry - XPC antibody [3.26] (ab6264)](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-2.jpg)
Overlay histogram showing A431 cells stained with ab6264 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6264, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
Western blot - Anti-XPC antibody [3.26] (ab6264)
![Western blot - Anti-XPC antibody [3.26] (ab6264)](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-3.jpg)
Predicted band size : 105 kDa
Western blot image of Human XPC from HeLa whole cell lysate using:
Lane 1: Rabbit polyclonal ab21078
Lane 2: Rabbit polyclonal ab21082
Lane 3: Rabbit monoclonal ab6264
This product has been referenced in:
See all 9 publications for this product
Publishing research using ab6264? Please let us know so that we can cite the reference in this datasheet
Concentration of lot no. is
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![Western blot - XPC antibody [3.26] (ab6264)](/ps/datasheet/Images/6/ab6264/ab6264_3.jpg)
Lane 1 : Anti-XPC antibody [3.26] (ab6264) at 1/1000 dilution
Lane 2 : Anti-XPC antibody [3.26] (ab6264) at 1/500 dilution
Lane 1 : Raji whole cell extract
Lane 2 : Raji whole cell extract
Predicted band size : 105 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
![Immunocytochemistry/ Immunofluorescence - XPC antibody [3.26] (ab6264)](/ps/datasheet/Images/6/ab6264/ab6264_4.jpg)
Ab6264 staining human XPC in HeLa cells (left), and DAPI staining (right) by immunofluorescence.
](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-1.jpg)
ab6264 (4µg/ml) staining XPC in human tonsil, using an automated system (DAKO Autostainer Plus). Using this protocol there is weak nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
![Flow Cytometry - XPC antibody [3.26] (ab6264)](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-2.jpg)
Overlay histogram showing A431 cells stained with ab6264 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6264, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (
![Western blot - Anti-XPC antibody [3.26] (ab6264)](/ps/datasheet/images/6/ab6264/XPC-Primary-antibodies-ab6264-3.jpg)
Predicted band size : 105 kDa
Western blot image of Human XPC from HeLa whole cell lysate using:
Lane 1: Rabbit polyclonal ab21078
Lane 2: Rabbit polyclonal ab21082
Lane 3: Rabbit monoclonal ab6264
1
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