Recombinant Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade KO Tested (ab259957)

Product name

Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade
See all RBBP7 primary antibodies
Description

Rabbit monoclonal [EPR23796-74] to RBBP7 - ChIP Grade
Host species

Rabbit
Tested applications

Suitable for: WB, IP, Flow Cyt (Intra), ChIP-sequencing, IHC-P, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HAP1, HeLa, HepG2, C6, Jurkat, MCF7, SH-SY5Y, LNCaP and F9 whole cell lysates; Mouse brain,heart, liver, and spleen tissue lysates; Rat heart, kidney and spleen tissue lysates. ICC/IF: HAP1, HeLa and NIH/3T3 cells. IHC-P: Human lung cancer tissue; Mouse lung tissue; Rat lung tissue. Flow Cyt (intra): HAP1, HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates. ChIP-Seq: Chromatin from HeLa cells.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23796-74
Isotype

IgG
Research areas

Alternative Versions
- Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade – BSA and Azide free (ab273882)
- Alexa Fluor® 488 Anti-RBBP7 antibody [EPR23796-74] (ab290984)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human RBBP7 (RbAp46) knockout HeLa cell line (ab264677)
KO cell lysates
- Human RBBP7 (RbAp46) knockout HeLa cell lysate (ab258628)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab259957 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
IP		1/30.
Flow Cyt (Intra)		1/600.
ChIP-sequencing		Use 3 µg for 30 µg of chromatin.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/500.

Notes
WB 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
IP 1/30.
Flow Cyt (Intra) 1/600.
ChIP-sequencing Use 3 µg for 30 µg of chromatin.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.

Function

Core histone-binding subunit that may target chromatin remodeling factors, histone acetyltransferases and histone deacetylases to their histone substrates in a manner that is regulated by nucleosomal DNA. Component of several complexes which regulate chromatin metabolism. These include the type B histone acetyltransferase (HAT) complex, which is required for chromatin assembly following DNA replication; the core histone deacetylase (HDAC) complex, which promotes histone deacetylation and consequent transcriptional repression; the nucleosome remodeling and histone deacetylase complex (the NuRD complex), which promotes transcriptional repression by histone deacetylation and nucleosome remodeling; and the PRC2/EED-EZH2 complex, which promotes repression of homeotic genes during development; and the NURF (nucleosome remodeling factor) complex.
Sequence similarities

Belongs to the WD repeat RBAP46/RBAP48/MSI1 family.
Contains 7 WD repeats.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q16576 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5931 Human
- Entrez Gene: 245688 Mouse
- Entrez Gene: 83712 Rat
- Omim: 300825 Human
- SwissProt: Q16576 Human
- SwissProt: Q60973 Mouse
- SwissProt: Q71UF4 Rat
- Unigene: 495755 Human
- Unigene: 270186 Mouse
- Unigene: 371732 Mouse
- Unigene: 3600 Rat
see all
Alternative names
- G1/S transition control protein binding protein RbAp46 antibody
- Histone acetyltransferase type B subunit 2 antibody
- Histone binding protein RBBP7 antibody
- Histone-binding protein RBBP7 antibody
- MGC138867 antibody
- MGC138868 antibody
- Nucleosome remodeling factor subunit RBAP46 antibody
- Nucleosome-remodeling factor subunit RBAP46 antibody
- RBAP46 antibody
- RBBP 7 antibody
- RBBP-7 antibody
- RBBP7 antibody
- RBBP7_HUMAN antibody
- Retinoblastoma binding protein 7 antibody
- Retinoblastoma binding protein p46 antibody
- Retinoblastoma-binding protein 7 antibody
- Retinoblastoma-binding protein p46 antibody
- Retinoblastoma-binding protein RbAp46 antibody
see all

ChIP-sequencing - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

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 ab259957 [EPR23796-74]. 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-sequencing - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ HeLa cells and 3 µg of ab259957 [EPR23796-74]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

At time of publication of this image, ChIP-seq was not widely characterised in HeLa for this antibody. For any questions, please contact Abcam Technical Support.

Additional screenshots of mapped reads can be downloaded here.
Western blot - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

All lanes : Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RBBP7 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 48 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Lanes 1 - 2: Merged signal (red and green). Green - ab259957 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab259957 was shown to react with RBBP7 in wild-type HeLa cells in Western blot with loss of signal observed in RBBP7 knockout cell line ab264677 (RBBP7 knockout cell lysate ab258628). Wild-type HeLa and RBBP7 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab259957 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RBBP7 (RbAp46) KO HAP1 cells labelling RBBP7 with ab259957 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing no staining in RBBP7 (RbAp46) KO HAP1 cells. 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 (2 ug/ml) dilution.
Western blot - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

All lanes : Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957) at 1/1000 dilution

Lane 1 : Jurkat (human t cell leukemia t lymphocyte) whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lane 4 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
Lane 5 : F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 48 kDa
Observed band size: 48 kDa

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

Exposure times: 8 seconds.
Western blot - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

All lanes : Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957) at 1/2000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : RBB7 (RbAp46) knockout HAP1 whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 48 kDa
Observed band size: 48 kDa

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

ab259957 was shown to specifically react with RBBP7 in wild-type HAP1 cells as signal was lost in RBB7 (RbAp46) knockout cells.

Wild-type and RBB7 (RbAp46) knockout samples were subjected to SDS-PAGE. ab259957 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/2000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc^™ MP instrument using the ECL technique.

Exposure time: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling RBBP7 with abab259957 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in mouse lung (PMID: 19655816). The section was incubated with ab259957 for 30 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling RBBP7 with ab259957 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing mainly nuclear staining in NIH/3T3 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 (2 ug/ml) dilution.
Flow Cytometry (Intracellular) - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized parental HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line (Right) / RBBP7 (RbAp46) KO HAP1 (Left) cells labelling RBBP7 with ab259957 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control 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.
Immunoprecipitation - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

RBBP7 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab259957 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259957 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2: abab259957 IP in NIH/3T3 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259957 in NIH/3T3 whole cell lysate

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

Exposure time: 24 seconds.
Immunoprecipitation - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

RBBP7 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab259957 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259957 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: abab259957 IP in HeLa whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259957 in HeLa whole cell lysate

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

Exposure time: 24 seconds.
Flow Cytometry (Intracellular) - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling RBBP7 with ab259957 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling RBBP7 with abab259957 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human lung cancer (PMID: 19655816).The section was incubated with ab259957 for 30 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry (Intracellular) - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Hela (Human cervix adenocarcinoma epithelial cell) cells labelling RBBP7 with ab259957 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control 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.
Immunocytochemistry/ Immunofluorescence - Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling RBBP7 with ab259957 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing mainly nuclear 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 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling RBBP7 with abab259957 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in rat lung (PMID: 19655816). The section was incubated with ab259957 for 30 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-RBBP7 antibody [EPR23796-74] (ab259957)

All lanes : Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Rat heart tissue lysate
Lane 6 : Rat kidney tissue lysate
Lane 7 : Rat spleen tissue lysate
Lane 8 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 9 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 10 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 11 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 48 kDa
Observed band size: 48 kDa

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

Exposure times: Lanes 1-3: 3 minutes; Lane 4: 3 seconds; Lanes 5-6: 3 minutes; Lane 7-11: 3 seconds.
Anti-RBBP7 antibody [EPR23796-74] (ab259957)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

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