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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-PKA R2/PKR2 抗体 [Y116] (ab32514)

Reviews (1) Submit a question References (8)
Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Flow Cytometry (Intracellular) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
  • Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y116] to PKA R2/PKR2
  • Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-PKA R2/PKR2 antibody [Y116]
    PKA R2/PKR2 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [Y116] to PKA R2/PKR2

  • 由来種

    Rabbit

  • 特異性

    The antibody recognises PKA RII. It does not cross-react with other cAMP-dependent kinase.

  • アプリケーション

    適用あり: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IFmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Cow

  • 免疫原

    Synthetic peptide within Human PKA R2/PKR2 aa 350-450 (C terminal). The exact sequence is proprietary.

  • ポジティブ･コントロール

    • WB: K562, HeLa and Jurkat cell lysates. IHC-P: Human colon cancer tissue. IP: K562 whole cell lysate. Flow Cyt (intra): MCF7 cells. ICC/IF: HeLa cells.

  • 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
    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

製品の特性

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ab32514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
Flow Cyt (Intra)
1/20.
IP
1/20.
WB (1)
1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 45 kDa).

For unpurified use at 1/10000 - 1/50000.
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.
ICC/IF
1/500.

For unpurified use at 1/100 - 1/250.
特記事項
Flow Cyt (Intra)
1/20.
IP
1/20.
WB
1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 45 kDa).

For unpurified use at 1/10000 - 1/50000.
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.
ICC/IF
1/500.

For unpurified use at 1/100 - 1/250.

ターゲット情報

  • 機能

    Regulatory subunit of the cAMP-dependent protein kinases involved in cAMP signaling in cells. Type II regulatory chains mediate membrane association by binding to anchoring proteins, including the MAP2 kinase.

  • 組織特異性

    Four types of regulatory chains are found: I-alpha, I-beta, II-alpha, and II-beta. Their expression varies among tissues and is in some cases constitutive and in others inducible.

  • 配列類似性

    Belongs to the cAMP-dependent kinase regulatory chain family.
    Contains 2 cyclic nucleotide-binding domains.

  • 翻訳後修飾

    Phosphorylated by the activated catalytic chain.

  • 細胞内局在

    Cytoplasm. Cell membrane. Colocalizes with PJA2 in the cytoplasm and the cell membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • cAMP dependent protein kinase regulatory subunit alpha 2 antibody
      • cAMP dependent protein kinase regulatory subunit RII alpha antibody
      • cAMP dependent protein kinase type II alpha regulatory chain antibody
      • cAMP dependent protein kinase type II alpha regulatory subunit antibody
      • cAMP-dependent protein kinase type II-alpha regulatory subunit antibody
      • KAP2 antibody
      • KAP2_HUMAN antibody
      • MGC3606 antibody
      • PKR 2 antibody
      • PKR2 antibody
      • PRKA R2 antibody
      • PRKAR 2 antibody
      • PRKAR2 antibody
      • PRKAR2A antibody
      • Protein kinase A RII alpha subunit antibody
      • Protein kinase cAMP dependent regulatory type II alpha antibody
      see all

    画像

    • Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/10000 dilution (Purified)

      Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 45 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

      Purified ab32514 at 1/20 dilution (0.5µg) immunoprecipitating PKA R2/PKR2 in K-562 whole cell lysate.
      Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
      Lane 2 (+): ab32514 + K-562 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32514 in K-562 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 50 kDa

    • Flow Cytometry (Intracellular) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Flow Cytometry (Intracellular) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

      Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/500 dilution (1.9 µg/mL). 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). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling PKA R2/PKR2 with purified ab32514 at 1/100 dilution (0.84 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Western blot - Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : PRKAR2A knockout HeLa cell lysate
      Lane 3 : K-562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 45 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab32514 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa. 

      Unpurified ab32514 Anti-PKA R2/PKR2 antibody [Y116] was shown to specifically react with PKA R2/PKR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265748 (knockout cell lysate ab257607) was used.  Wild-type and PKA R2/PKR2 knockout samples were subjected to SDS-PAGE.  ab32514 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 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 20000 dilution for 1 hour at room temperature before imaging.

    • Anti-PKA R2/PKR2 antibody [Y116] (ab32514)
      Anti-PKA R2/PKR2 antibody [Y116] (ab32514)

    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.

    Click here to view the general protocols

    参考文献 (8)

    ab32514 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab32514 は 8 報の論文で使用されています。

    • Jiang Y  et al. Type IIB PKA is highly expressed in β cells and controls cell proliferation via regulating Cyclin D1 expression. FEBS J 289:2865-2876 (2022). PubMed: 34839588
    • Peng L  et al. Prostaglandin E2 Receptor 4 (EP4) Affects Trophoblast Functions via Activating the cAMP-PKA-pCREB Signaling Pathway at the Maternal-Fetal Interface in Unexplained Recurrent Miscarriage. Int J Mol Sci 22:N/A (2021). PubMed: 34502044
    • Xie S  et al. miR-1307 promotes hepatocarcinogenesis by CALR-OSTC-endoplasmic reticulum protein folding pathway. iScience 24:103271 (2021). PubMed: 34761190
    • Hedderich J  et al. Norepinephrine Inhibits the Proliferation of Human Bone Marrow-Derived Mesenchymal Stem Cells via ß2-Adrenoceptor-Mediated ERK1/2 and PKA Phosphorylation. Int J Mol Sci 21:N/A (2020). PubMed: 32486305
    • Speichert S  et al. Role of Norepinephrine in IL-1ß-Induced Chondrocyte Dedifferentiation under Physioxia. Int J Mol Sci 20:N/A (2019). PubMed: 30861996
    • El Bagdadi K  et al. Norepinephrine Inhibits Synovial Adipose Stem Cell Chondrogenesis via a2a-Adrenoceptor-Mediated ERK1/2 Activation. Int J Mol Sci 20:N/A (2019). PubMed: 31248037
    • Hillman P  et al. Dissociation between AKAP3 and PKARII promotes AKAP3 degradation in sperm capacitation. PLoS One 8:e68873 (2013). WB ; Cow . PubMed: 23894359
    • Gu L  et al. Protein kinase A activation confers resistance to trastuzumab in human breast cancer cell lines. Clin Cancer Res 15:7196-206 (2009). IHC-P ; Human . PubMed: 19920112

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    Western blot abreview for Anti-PKA R2/PKR2 antibody [Y116]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Caudate)
    Gel Running Conditions
    Reduced Denaturing (NW04127 17 well gel, ThermoFisher))
    Loading amount
    20 µg
    Specification
    Caudate
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Antón Leandro Martínez Rodríguez

    Verified customer

    投稿 Aug 02 2022

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