Anti-NF-kB p65 (acetyl K310)抗体

• IP

Unknown

Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (AB19870)

Western Blot with ab19870 after p65 Immunoprecipitation : rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5μg/ml) in 1% non-fat milk TBS-T incubated for 3 hours at room temperature. Exposure time : 1 min normal ECL. Tested samples : nuclear extracts (180 μg) of immortalized p65-/- mouse cells, complemented with the empty vector (pRRL), wild-type p65 (Wt) and non-acetylatable K310 (K310R). The samples tested were treated with deacetylase inhibitors HDACi (TSA + Nicotinamide) and TNF-alpha. The samples were immunoprecipitated with 2μg of alpha-p65 and subsequently analysed by Western blot with Rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870). Predicted band size = 65kDa, Observed band size = 75kDa. The p65 band runs higher in this SDS-PAGE blot as it contains a myc-tag.

All lanes:

Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)

Predicted band size: 60 kDa

false

This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland

• WB

Characteriser7094120****

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (AB19870)

Observed band size : 75kDa

The p65 band runs higher in this blot because the protein contains a myc-tag.

Lanes 1 - 2:

Coomassie stain

Lanes 3 - 4:

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1/500 dilution

Lanes 1 and 3:

Acetylated p65 protein at 2 µg

Lanes 2 and 4:

K310R mutant protein at 2 µg

Secondary

All lanes:

IRDye® 800CW Goat anti-Rabbit IgG (H + L) at 1/15000 dilution

Predicted band size: 60 kDa

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This image is courtesy of Prof. Michael Hottiger's Lab, University of Zurich

• WB

Unknown

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (AB19870)

ab19870 recognizes Rabbit polyclonal to NF-kB p65 (acetyl K310) specifically at ~75kDa (indicated by the arrow) is this SDS-PAGE blot. The p65 band runs higher than 65kDa in this SDS-PAGE blot as it contains a myc-tag. We are sure that the band at ~75kDa is p65 since p65 specific antibodies detect the same band in IP and WB and there is no signal in the p65 knock-out cell line with ab19870. A number of additonal bands are recognized by ab19870 when tested with endogenous p65 from whole cell extracts, we do not know the identity of these bands.

Tested samples : nuclear extracts (75μg) of immortalized p65-/- mouse cells, complemented with the empty ve

All lanes:

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 2.5 µg/mL

Lane 1:

pRRL untreated at 75 µg

Lane 2:

pRRL HDACi at 75 µg

Lane 3:

pRRL HDACi + TNF at 75 µg

Lane 4:

Wt untreated at 75 µg

Lane 5:

Wt HDACi at 75 µg

Lane 6:

Wt HDACi + phorbol myristate acetate at 75 µg

Lane 7:

K310R untreated at 75 µg

Lane 8:

K310R HDACi at 75 µg

Lane 9:

K310R HDACi + phorbol myristate acetate at 75 µg

Predicted band size: 60 kDa

Observed band size: 75 kDa

true

Exposure time: 1hr

This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland

• WB

Ap876607****

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (AB19870)

All lanes:

Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1 µg/mL

All lanes:

Lung (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 60 kDa

Observed band size: 15 kDa,72 kDa

false

Exposure time: 4min

• Dot

Collaborator

Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (AB19870)

Rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5μg/ml) in 1% non-fat milk TBS-T incubated for 3h at room temperature. Exposure time : 75 min normal ECL. This Dot blot demonstrates that ab19870 recognized upto 10ng of purified peptide on a PVDF membrane.

This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland