Anti-SMARCC1/BAF155 抗体 [EPR12395] - ChIP Grade (ab172638)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12395] to SMARCC1/BAF155 - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, WB, ICC/IF, IP
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-SMARCC1/BAF155 antibody [EPR12395] - ChIP Grade
SMARCC1/BAF155 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR12395] to SMARCC1/BAF155 - ChIP Grade -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), ChIP, WB, ICC/IF, IPmore details -
種交差性
交差種: Rat, Human -
免疫原
Synthetic peptide within Human SMARCC1/BAF155 aa 700-800 (Cysteine residue). The exact sequence is proprietary.
Database link: Q92922 -
ポジティブ・コントロール
- HeLa, K562, Jurkat and 293T cell lysates. Permeabilized Jurkat cells. Immunoprecipitation pellet from Jurkat whole cell lysate (ab7899). Rat testis lysates.
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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.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.21% 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
EPR12395 -
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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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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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ab172638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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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIP |
Use at an assay dependent concentration.
|
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 123 kDa.
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ICC/IF | (1) |
1/100.
For unpurified use at 1/250 - 1/500. |
IP |
1/10 - 1/100.
|
特記事項 |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Predicted molecular weight: 123 kDa. |
ICC/IF
1/100. For unpurified use at 1/250 - 1/500. |
IP
1/10 - 1/100. |
ターゲット情報
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機能
Involved in transcriptional activation and repression of select genes by chromatin remodeling (alteration of DNA-nucleosome topology). May stimulate the ATPase activity of the catalytic subunit of the complex. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene. Belongs to the neural progenitors-specific chromatin remodeling complex (npBAF complex) and the neuron-specific chromatin remodeling complex (nBAF complex). During neural development a switch from a stem/progenitor to a post-mitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state. The transition from proliferating neural stem/progenitor cells to post-mitotic neurons requires a switch in subunit composition of the npBAF and nBAF complexes. As neural progenitors exit mitosis and differentiate into neurons, npBAF complexes which contain ACTL6A/BAF53A and PHF10/BAF45A, are exchanged for homologous alternative ACTL6B/BAF53B and DPF1/BAF45B or DPF3/BAF45C subunits in neuron-specific complexes (nBAF). The npBAF complex is essential for the self-renewal/proliferative capacity of the multipotent neural stem cells. The nBAF complex along with CREST plays a role regulating the activity of genes essential for dendrite growth. -
組織特異性
Expressed in brain, heart, muscle, placenta, lung, liver, muscle, kidney and pancreas. -
配列類似性
Belongs to the SMARCC family.
Contains 1 SANT domain.
Contains 1 SWIRM domain. -
翻訳後修飾
Phosphorylated on undefined residues at the G2/M transition by ERK1 and other kinases. This may contribute to cell cycle specific inactivation of remodeling complexes containing the phosphorylated protein. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 6599 Human
- Entrez Gene: 301020 Rat
- Omim: 601732 Human
- SwissProt: Q92922 Human
- Unigene: 476179 Human
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別名
- AI115498 antibody
- BAF 155 antibody
- BAF155 antibody
see all
画像
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All lanes : Anti-SMARCC1/BAF155 antibody [EPR12395] - ChIP Grade (ab172638) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SMARCC1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 123 kDa
Observed band size: 112 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab172638 observed at 112 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab172638 was shown to react with SMARCC1 in wild-type HeLa cells in Western blot with loss of signal observed in SMARCC1 knockout cell line ab264859 (SMARCC1 knockout cell lysate ab258198). Wild-type HeLa and SMARCC1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab172638 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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All lanes : Anti-SMARCC1/BAF155 antibody [EPR12395] - ChIP Grade (ab172638) at 1/5000 dilution
Lane 1 : Wild-type HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : SMARCC1 knockout HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 123 kDaLanes 1 - 4: Merged signal (red and green). Green - ab172638 observed at 123 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab172638 was shown to recognize in wild-type HEK293 cells as signal was lost at the expected MW in SMARCC1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SMARCC1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab172638 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SMARCC2 with Purified ab172638 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor ® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor ® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling SMARCC1/BAF155 with purified ab172638 at 1/30 dilution (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).
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Chromatin was prepared from MDA-MB-231 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab172638 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci).
Primers and probes are located in the first kb of the transcribed region. -
All lanes : Anti-SMARCC1/BAF155 antibody [EPR12395] - ChIP Grade (ab172638) at 1/5000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Rat testis lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 123 kDa
Observed band size: 155 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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ab172638 (purified) at 1:30 dilution (2μg) immunoprecipitating SMARCC1/BAF155 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate, 10μg
Lane 2 (+): ab172638 & Jurkat whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab172638 in Jurkat 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." -
Western blot analysis on immunoprecipitation pellet from Jurkat cell lysate using unpurified ab172638 at a 1/10 dilution.
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Intracellular flow cytometric analysis of permeabilized Jurkat cells usingunpurified ab172638 at a 1/10 dilution (red) or a rabbit IgG (negative) (green).
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All lanes : Anti-SMARCC1/BAF155 antibody [EPR12395] - ChIP Grade (ab172638) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : K562 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 123 kDa
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (6)
ab172638 は 6 報の論文で使用されています。
- Wei Z et al. SMARCC1 Enters the Nucleus via KPNA2 and Plays an Oncogenic Role in Bladder Cancer. Front Mol Biosci 9:902220 (2022). PubMed: 35669562
- Wu S et al. Targeting glutamine dependence through GLS1 inhibition suppresses ARID1A-inactivated clear cell ovarian carcinoma. Nat Cancer 2:189-200 (2021). PubMed: 34085048
- Cyrta J et al. Role of specialized composition of SWI/SNF complexes in prostate cancer lineage plasticity. Nat Commun 11:5549 (2020). IP ; Human . PubMed: 33144576
- Wu S et al. ARID1A spatially partitions interphase chromosomes. Sci Adv 5:eaaw5294 (2019). PubMed: 31131328
- Scott CA et al. Nuclear/cytoplasmic transport defects in BBS6 underlie congenital heart disease through perturbation of a chromatin remodeling protein. PLoS Genet 13:e1006936 (2017). PubMed: 28753627
- Cabot B et al. Differential expression of key subunits of SWI/SNF chromatin remodeling complexes in porcine embryos derived in vitro or in vivo. Mol Reprod Dev 84:1238-1249 (2017). ICC/IF . PubMed: 29024220