Anti-RBBP7 抗体 [EPR23796-74] - ChIP Grade (ab259957)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23796-74] to RBBP7 - ChIP Grade
- Suitable for: WB, IP, Flow Cyt (Intra), ChIP-sequencing, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade
RBBP7 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR23796-74] to RBBP7 - ChIP Grade -
由来種
Rabbit -
アプリケーション
適用あり: WB, IP, Flow Cyt (Intra), ChIP-sequencing, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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.
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特記事項
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR23796-74 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab259957の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
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IP |
1/30.
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Flow Cyt (Intra) |
1/600.
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ChIP-sequencing |
Use 3 µg for 30 µg of chromatin.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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特記事項 |
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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. |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the WD repeat RBAP46/RBAP48/MSI1 family.
Contains 7 WD repeats. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- 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
see all -
別名
- G1/S transition control protein binding protein RbAp46 antibody
- Histone acetyltransferase type B subunit 2 antibody
- Histone binding protein RBBP7 antibody
see all
画像
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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.
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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.
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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.
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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.
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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 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: 8 seconds.
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All lanes : Anti-RBBP7 antibody [EPR23796-74] - ChIP Grade (ab259957) at 1/2000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : RBBP7 (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 kDaBlocking 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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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 kDaBlocking 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.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (0)
ab259957 は論文での使用が確認できていません。