Recombinant Anti-RBBP4 antibody [EPR3411] - ChIP Grade (ab79416)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8 µg of ab79416 [EPR3411]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

Additional screenshots of mapped reads can be downloaded here.

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 3 µg of ab79416. ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

Additional screenshots of mapped reads can be downloaded here.

DOI: 10.7554/eLife.54993.

Blocking and diluting buffer: 5% NFDM/TBST. 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat pancreas tissue sections labeling RBBP4 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

ab79416 (purified) at 1:20 dilution (2µg) immunoprecipitating RBBP4 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab79416 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab79416 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling RBBP4 with purified ab79416 at 1/20 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling RBBP4 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling RBBP4 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylinwas used as a counterstain

Blocking and diluting buffer: 5% NFDM/TBST. 

ab79416 staining RBBP4 in MCF-7 (human breast carcinoma) cells by ICC (Immunocytochemistry). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

Negative control 1: PBS only.

Unpurifed ab79416, at 1/100 dilution, staining RBBP4 in human breast carcinoma tissue by Immunohistochemistry.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

Immunocytochemical staining of RBBP4 in HeLa cells using unpurified ab79416 at 1/100 dilution.