Anti-hnRNP A2B1 抗体 (ab31645)
Key features and details
- Rabbit polyclonal to hnRNP A2B1
- Suitable for: IHC-P, WB, ICC, IP
- Reacts with: Human
- Isotype: IgG
製品の概要
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製品名
Anti-hnRNP A2B1 antibody
hnRNP A2B1 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to hnRNP A2B1 -
由来種
Rabbit -
特異性
The peptide used as the immunogen for this antibody is found within isoform A2 and B1 of the protein and should thus recognize a doublet around 35/37 kDa. From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
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アプリケーション
適用あり: IHC-P, WB, ICC, IPmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat, Chicken, Dog, Xenopus laevis -
免疫原
Synthetic peptide corresponding to Human hnRNP A2B1 aa 50-150.
(Peptide available asab31644) -
ポジティブ・コントロール
- WB: A549, HEK293T, HeLa, NIH 3T3, and MEF1 cell lysates, and mouse testis tissue lysate. IHC-P: Human colon tissue. ICC: HeLa cells.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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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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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ab31645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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IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB | (2) |
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 35, 38 kDa (predicted molecular weight: 35 , 37 kDa).
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ICC |
Use a concentration of 5 µg/ml.
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IP |
Use at an assay dependent concentration.
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特記事項 |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 35, 38 kDa (predicted molecular weight: 35 , 37 kDa). |
ICC
Use a concentration of 5 µg/ml. |
IP
Use at an assay dependent concentration. |
ターゲット情報
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機能
Involved with pre-mRNA processing. Forms complexes (ribonucleosomes) with at least 20 other different hnRNP and heterogeneous nuclear RNA in the nucleous. -
配列類似性
Contains 2 RRM (RNA recognition motif) domains. -
細胞内局在
Nucleus > nucleoplasm. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Predominantly nucleoplasmic, however isoform A2 is also found in the cytoplasm of cells in some tissues. Not found in the nucleolus. - Information by UniProt
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参照データベース
- Entrez Gene: 3181 Human
- Entrez Gene: 100045191 Mouse
- Entrez Gene: 53379 Mouse
- Entrez Gene: 362361 Rat
- Entrez Gene: 380082 Xenopus laevis
- Omim: 600124 Human
- SwissProt: P22626 Human
- SwissProt: O88569 Mouse
see all -
別名
- Heterogeneous nuclear ribonucleoprotein A2 antibody
- Heterogeneous nuclear ribonucleoprotein A2/B1 antibody
- Heterogeneous nuclear ribonucleoprotein B1 antibody
see all
画像
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All lanes : Anti-hnRNP A2B1 antibody (ab31645) at 1/500 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : HNRNPA2B1 knockout HEK293T cell lysate
Lane 3 : A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 , 37 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab31645 observed at 37 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab31645 Anti-hnRNP A2B1 antibody was shown to specifically react with hnRNP A2B1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266404 (knockout cell lysate ab257224) was used. Wild-type and hnRNP A2B1 knockout samples were subjected to SDS-PAGE. ab31645 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti- Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti- Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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ab31645 staining hnRNP A2B1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 overnight at 4°C with ab31645 at 5 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Anti-hnRNP A2B1 antibody (ab31645) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 35 , 37 kDa
Observed band size: 35,38 kDa why is the actual band size different from the predicted?
ab31645 is expected to recognize isoform hnrnp A2 and hnrnp B1. The antibody detects bands at approximately 35 and 38 kDa which are of the correct size to correspond to the predicted molecular weight of these isoforms. -
hnRNP A2B1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to hnRNP A2B1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31645.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 35kDa: hnRNP A2B1. -
IHC image of ab31645 staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31645, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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 (21)
ab31645 は 21 報の論文で使用されています。
- Wheeler JR et al. RNA-binding proteins direct myogenic cell fate decisions. Elife 11:N/A (2022). PubMed: 35695839
- Asamitsu S et al. CGG repeat RNA G-quadruplexes interact with FMRpolyG to cause neuronal dysfunction in fragile X-related tremor/ataxia syndrome. Sci Adv 7:N/A (2021). PubMed: 33523882
- Luo J et al. lncRNA PCAT6 facilitates cell proliferation and invasion via regulating the miR-326/hnRNPA2B1 axis in liver cancer. Oncol Lett 21:471 (2021). PubMed: 33907581
- Liu B et al. Enzalutamide-Induced Upregulation of PCAT6 Promotes Prostate Cancer Neuroendocrine Differentiation by Regulating miR-326/HNRNPA2B1 Axis. Front Oncol 11:650054 (2021). PubMed: 34277403
- Lambert M et al. Identification of Abundant and Functional dodecaRNAs (doRNAs) Derived from Ribosomal RNA. Int J Mol Sci 22:N/A (2021). PubMed: 34575920