Anti-RBBP4 antibody [EPR3411] - ChIP Grade

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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).

• IP

Unknown

Immunoprecipitation - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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.

All lanes:

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

Predicted band size: 48 kDa

Observed band size: 33 kDa,42 kDa

false

• ChIP-seq

Supplier Data

ChIP-sequencing - 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.

• ChIP-seq

Supplier Data

ChIP-sequencing - 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 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

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

• WB

Unknown

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (ab79416) at 0.006 µg/mL

All lanes:

Human fetal spleen lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 48 kDa

false

• WB

Unknown

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (ab79416) at 0.006 µg/mL

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

Mouse spleen lysates at 15 µg

Lane 3:

Rat spleen lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 48 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab79416 [EPR3411]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab79416 [EPR3411]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab79416 [EPR3411]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Unknown

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (AB79416)

All lanes:

Western blot - Anti-RBBP4 antibody [EPR3411] - ChIP Grade (ab79416) at 1/50000 dilution

Lane 1:

HeLa lysate at 10 µg

Lane 2:

Jurkat lysate at 10 µg

Lane 3:

NIIH/3T3 lysate at 10 µg

Lane 4:

293 lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

false