Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling KAT3B/p300+CREBBP with ab275379 at 1/5000 (0.093 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human hepatocellular carcinoma (PMID: 32144235) is observed. The section was incubated with ab275379 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling KAT3B/p300+CREBBP with ab275379 at 1/100 (4.67 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000  dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling KAT3B/p300+CREBBP with ab275379 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

All lanes : Anti-CREBBP + KAT3B / p300 antibody [EPR23495-276] (ab275379) at 1/1000 dilution

Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Observed band size: 300 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID: 8995708).

Lysates were made freshly and used in WB immediately to minimize protein degradation.

Exposure times: Lane 1: 48 seconds; Lane 2: 70 seconds; Lane 3: 59 seconds.

 

Immunohistochemical analysis of paraffin-embedded Human clear cell renal cell carcinoma tissue labeling KAT3B/p300+CREBBP with ab275379 at 1/5000 (0.093 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human clear cell renal cell carcinoma (PMID: 28551630). The section was incubated with ab275379 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

All lanes : Anti-CREBBP + KAT3B / p300 antibody [EPR23495-276] (ab275379) at 1/1000 dilution

Lane 1 : His-tagged human KAT3B/p300 recombinant protein, 20 ng
Lane 2 : His-tagged human CREBBP recombinant protein, E. coli extract, 10 µl

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

This product can recognize both KAT3B/p300 and CREBBP.

Exposure time: 3 seconds.

 

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling KAT3B/p300+CREBBP with ab275379 at 1/5000 (0.093 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human colon (PMID: 28586030). The section was incubated with ab275379 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Anti-CREBBP + KAT3B / p300 antibody [EPR23495-276] (ab275379) at 1/1000 dilution + HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary
VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/5000 dilution

Observed band size: 300 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID: 8995708).

Lysate was made freshly and used in WB immediately to minimize protein degradation.

Exposure time: 32 seconds.

 

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling KAT3B/p300+CREBBP with ab275379 at 1/5000 (0.093 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human kidney (PMID: 28551630). The section was incubated with ab275379 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.