Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5248] to KAP1 (phospho S824)
- Suitable for: WB, IP, Dot blot
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-KAP1 (phospho S824) antibody [EPR5248]
See all KAP1 primary antibodies -
Description
Rabbit monoclonal [EPR5248] to KAP1 (phospho S824) -
Host species
Rabbit -
Specificity
ab133440 only detects KAP1 phosphorylated at Serine 824. -
Tested Applications & Species
See all applications and species dataApplication Species IP HumanWB MouseHuman -
Immunogen
Synthetic phospho peptide corresponding to a region surrounding Serine 824 of Human KAP1 (UniProt Q13263).
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Positive control
- Lysate from HeLa cells treated with gamma radiation. IP: HeLa treated with 3uM etoposide for 1h whole cell lysate.
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR5248 -
Isotype
IgG -
Research areas
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Wild type HAP1 + DMSO whole cell lysate (20 µg)
Lane 3: Wild type HAP1 + Bleomycin whole cell lysate (20 µg)
Lane 4: KAP1 knockout HAP1 whole cell lysate (20 µg)
Lane 5: KAP1 knockout HAP1 + DMSO whole cell lysate (20 µg)
Lane 6: KAP1 knockout HAP1 + Bleomycin whole cell lysate (20 µg)
Lane 7: HeLa + DMSO whole cell lysate (20 µg)
Lane 8: HeLa + Bleomycin whole cell lysate (20 µg)Lanes 1 - 8: Merged signal (red and green). Green - ab133440 observed at 105 kDa. Red - loading control, ab130007, observed at 125 kDa.
ab133440 was shown to specifically react with KAP1 in wild type cells as signal was lost in KAP1 knockout cells. Wild-type and KAP1 knockout samples were subjected to SDS-PAGE. ab133440 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C both at a 1/20000 dilution. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. Treated with 30 µg/mL Bleomycin in DMSO for 30 minutes.
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Purified ab133440 at 1/50 dilution (2µg) immunoprecipitating KAP1 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with 3uM etoposide for 1h whole cell lysate 10µg
Lane 2 (+): ab133440 + HeLa treated with 3uM etoposide for 1h whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32132 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 110 kDa -
All lanes : Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 3µM etoposide for 1h whole cell lysate
Lane 3 : HeLa treated with 3µM etoposide for 1h whole cell lysate. Then the membrane was incubated with alkaline phosphatase.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440) at 1/1000 dilution
Lane 1 : MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : MDA-MB-231 treated with 1µM camptothecin for 24h whole cell lysate
Lane 3 : MDA-MB-231 treated with 1µM camptothecin for 24h whole cell lysate. Then the membrane was incubated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of KAP1 (phospho S824) phospho peptide (Lane 1) and KAP1 non-phospho peptide (Lane 2) labelling KAP1 (phospho S824) with ab133440 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.
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All lanes : Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440) at 1/50000 dilution
Lane 1 : Lysate of HeLa cells, untreated
Lane 2 : Lysate of HeLa cells treated with gamma radiation
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit at 1/2000 dilution
Predicted band size: 88 kDa
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