Antibodies, Proteins, Kits and Reagents for Life Science

328 339 ₸ Product size

100 µl 328 339 ₸ 10 µl 134 016 ₸

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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

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

Application Species
IP
Human

WB
Mouse

Human

See all applications and species data
Immunogen

Synthetic phospho peptide corresponding to a region surrounding Serine 824 of Human KAP1 (UniProt Q13263).
Positive control
- Lysate from HeLa cells treated with gamma radiation. IP: HeLa treated with 3uM etoposide for 1h whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Application	Species
IP	Human
WB	Mouse Human

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...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR5248
Isotype

IgG
Research areas
- Cancer
- Cell Death
- Apoptosis
- Other
- Epigenetics and Nuclear Signaling
- Bromodomains

Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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.
Immunoprecipitation - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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
Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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.
Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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.
Dot Blot - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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.
Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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
Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440)

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