Anti-XPA antibody [5F12] (ab65963)
Key features and details
- Mouse monoclonal [5F12] to XPA
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG2b
Overview
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Product name
Anti-XPA antibody [5F12]
See all XPA primary antibodies -
Description
Mouse monoclonal [5F12] to XPA -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human -
Immunogen
Recombinant full length XPA protein (Human)
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Epitope
Amino acids 30-47. -
Positive control
- WB: HeLa and 293T cell lysates.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 6
Constituents: PBS, 50% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Ion Exchange Chromatography -
Clonality
Monoclonal -
Clone number
5F12 -
Isotype
IgG2b -
Research areas
Images
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All lanes : Anti-XPA antibody [5F12] (ab65963) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : XPA knockout HeLa cell lysate
Lane 3 : 293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?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.
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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.
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Anti-XPA antibody [5F12] (ab65963) at 0.1 µg/ml + Crude extract of HeLa cells at 200 µg
Predicted band size: 31 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
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All lanes : 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