Anti-ARID1A 抗体 [EPR13501] - BSA and Azide free (ab217154)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13501] to ARID1A - BSA and Azide free
- Suitable for: ICC/IF, ChIC/CUT&RUN-seq, Flow Cyt (Intra), IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-ARID1A antibody [EPR13501] - BSA and Azide free
ARID1A 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR13501] to ARID1A - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, ChIC/CUT&RUN-seq, Flow Cyt (Intra), IHC-P, WBmore details
適用なし: ChIP -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat -
免疫原
Recombinant fragment. 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
- IHC-P: Human kidney and human adenocarcinoma of endometrium without ARID1A mutation tissues. ICC/IF: Wildtype HAP1 and SH-SY5Y cells. WB: HEK-293T and SH-SY5Y cell lysates. ChIC/CUT&RUN-Seq: HCT116 cells.
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特記事項
ab217154 is the carrier-free version of ab182560.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
バッファー
pH: 7.20
Constituent: PBS -
キャリア・フリー
はい -
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
EPR13501 -
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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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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ab217154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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ICC/IF |
Use at an assay dependent concentration.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 242 kDa.
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特記事項 |
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ICC/IF
Use at an assay dependent concentration. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 242 kDa. |
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). Binds DNA non-specifically. 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
Highly expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon, and PBL, and at a much lower level in heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. -
配列類似性
Contains 1 ARID domain. -
翻訳後修飾
Phosphorylated upon DNA damage, probably by ATM or ATR. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 8289 Human
- Entrez Gene: 93760 Mouse
- Entrez Gene: 297867 Rat
- Omim: 603024 Human
- SwissProt: O14497 Human
- SwissProt: A2BH40 Mouse
- Unigene: 468972 Human
- Unigene: 22478 Mouse
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別名
- actin-dependent regulator of chromatin subfamily F member 1 antibody
- ARI1A_HUMAN antibody
- ARID domain containing protein 1A antibody
see all
画像
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HCT116 cells and 5µg of ab182560 [EPR13501]. 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.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
This data was developed using the same antibody clone in a different buffer formulation (ab182560).
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All lanes : Anti-ARID1A antibody [EPR13501] (ab182560) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : ARID1A knockout HEK-293T cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 242 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab182560).
Lanes 1 - 3: Merged signal (red and green). Green - ab182560 observed at 270 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab182560 was shown to react with ARID1A in wild-type HEK-293T cells in Western blot with loss of signal observed in ARID1A knockout cell line ab266189 (ARID1A knockout cell lysate ab257250). Wild-type HEK-293T and ARID1A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab182560 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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Immunohistochemical analysis of paraffin embedded Human adenocarcinoma of endometrium without ARID1A mutation (Left image) labeling ARID1A using ab182560 at 1/1000 dilution. Right image: Right picture: paraffine embedded human adenocarcinoma of endometrium with ARID1A mutation. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling ARID1A with purified ab182560 at 1/230 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).
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Anti-ARID1A antibody [EPR13501] (ab182560) at 1/1000 dilution + 293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 242 kDa
Observed band size: 130,270 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsThis data was developed using the same antibody clone in a different buffer formulation (ab182560).Blocking buffer and concentration: 5% NFDM/TBSObserved band: 130-270 kDaARID1A has many mutations which typically generate truncated proteins that are highly prone to degradation. (PMID: 21614196, PMID: 29486633, PMID: 34429326). -
ab182560 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab182560 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).
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This IHC data was generated using the same anti-ARID1A antibody clone, EPR13501, in a different buffer formulation (cat# ab182560).
Immunohistochemical analysis of paraffin embedded Human kidney tissue (Left image) labeling ARID1A using ab182560 at 1/1000 dilution. Right image: Right picture: paraffine embedded human clear cell carcinoma of kidney with ARID1A mutation. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Clone EPR13501 (ab217154) has been successfully conjugated by Abcam. This image was generated using Anti-ARID1A antibody [EPR13501] (Alexa Fluor® 647). Please refer to ab216304 for protocol details.
ab216304 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab216304 at 1/500 dilution (shown in red) and ab195887 at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone EPR13501 (ab217154) has been successfully conjugated by Abcam. This image was generated using Anti-ARID1A antibody [EPR13501] (Alexa Fluor® 488). Please refer to ab216112 for protocol details.
ab216112 staining ARID1A in wild-type HAP1 cells (top panel) and ARID1A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab216112 at 1/500 dilution (shown in green) and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunofluorescent analysis of SH-SY5Y cells labeling ARID1A with ab182560 at 1/500 and Goat anti rabbit IgG(Alexa Fluor®555) at 1/200. Image at the right stained with DAPI.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182560).
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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Datasheet download
Certificate of Compliance
参考文献 (1)
ab217154 は 1 報の論文で使用されています。
- Concepcion CP et al. Smarca4 Inactivation Promotes Lineage-Specific Transformation and Early Metastatic Features in the Lung. Cancer Discov 12:562-585 (2022). PubMed: 34561242