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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-PINK1 抗体 [MJF-R32-7] - BSA and Azide free (ab300624)

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Western blot - Anti-PINK1 antibody [MJF-R32-7] - BSA and Azide free (ab300624)
  • Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (ab300624)
  • Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)
  • Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJF-R32-7] to PINK1 - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

製品の概要

  • 製品名

    Anti-PINK1 antibody [MJF-R32-7] - BSA and Azide free
    PINK1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [MJF-R32-7] to PINK1 - BSA and Azide free

  • 由来種

    Rabbit

  • 特異性

    This antibody does not react in: IHC-P with human, mouse and rat; and in immunocytochemistry, flow cytometry and immunoprecipitation with human.

    This antibody was mapped to AA 188-194 with some cross-reaction to AA 287-301.

  • アプリケーション

    適用あり: WBmore details
    適用なし: Flow Cyt,ICC/IF,IHC-P or IP

  • 種交差性

    交差種: Human
    非交差種: Mouse, Rat

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Wild-type HEK-293T treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 6 hours, whole cell lysate; Untreated PC-3 whole cell lysate; PC-3 treated with CCCP for 6 hours, whole cell lysate.

  • 特記事項

    ab30064 is a carrier free version of ab300623. 

     

    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
    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.

    This antibody was developed with support from The Michael J. Fox Foundation.

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ab300624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
WB
Use at an assay dependent concentration. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).
特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).
追加情報
Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.

ターゲット情報

  • 機能

    Protects against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondrial proteins. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). It is necessary for PARK2 recruitement to dysfunctional mitochondria to initiate their degradation.

  • 組織特異性

    Highly expressed in heart, skeletal muscle and testis, and at lower levels in brain, placenta, liver, kidney, pancreas, prostate, ovary and small intestine. Present in the embryonic testis from an early stage of development.

  • 関連疾患

    Defects in PINK1 are the cause of Parkinson disease type 6 (PARK6) [MIM:605909]. A neurodegenerative disorder characterized by parkinsonian signs such as rigidity, resting tremor and bradykinesia. A subset of patients manifest additional symptoms including hyperreflexia, autonomic instability, dementia and psychiatric disturbances. Symptoms show diurnal fluctuation and can improve after sleep.

  • 配列類似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
    Contains 1 protein kinase domain.

  • 翻訳後修飾

    Autophosphorylated.

  • 細胞内局在

    Mitochondrion outer membrane. Cytoplasm > cytosol.
  • Information by UniProt

  • 参照データベース

    • 別名

      • BRPK antibody
      • FLJ27236 antibody
      • mitochondrial antibody
      • PARK 6 antibody
      • PARK6 antibody
      • Phosphatase and Tensin Homolog antibody
      • PINK 1 antibody
      • PINK1 antibody
      • PINK1_HUMAN antibody
      • Protein kinase BRPK antibody
      • PTEN induced putative kinase 1 antibody
      • PTEN induced putative kinase protein 1 antibody
      • PTEN-induced putative kinase protein 1 antibody
      • Serine/threonine kinase PINK1 mitochondrial antibody
      • Serine/threonine protein kinase PINK1 mitochondrial antibody
      • Serine/threonine-protein kinase PINK1 antibody
      see all

    画像

    • Western blot - Anti-PINK1 antibody [MJF-R32-7] - BSA and Azide free (ab300624)
      Western blot - Anti-PINK1 antibody [MJF-R32-7] - BSA and Azide free (ab300624)
      All lanes : Anti-PINK1 antibody [MJF-R32-7] (ab300623) at 1/1000 dilution

      Lane 1 : Wild-type SH-SY5Y treated CCCP, ab141229 (10 µM, 24 h) cell lysate
      Lane 2 : Wild-type SH-SY5Y control CCCP (0 uM, 24 h) cell lysate
      Lane 3 : PINK1 knockout SH-SY5Y (ab280876) treated CCCP, ab141229 (10 uM, 24 h) cell lysate
      Lane 4 : PINK1 SH-SY5Y (ab280876) control CCCP (0 uM, 24 h) cell lysate
      Lane 5 : Wild-type HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate
      Lane 6 : PINK1 knockout HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 63 kDa
      Observed band size: 63 kDa



      This data was developed using ab300623, the same antibody clone in a different buffer formulation. 

      Western blot: Anti-PINK1 antibody [MJF-R32-7] (ab300623) staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta.

      In Western blot, ab300623 was shown to bind specifically to PINK1. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

      In Western blot, ab300623 was shown to bind specifically to PINK1. A band was observed at 63kDa in wild-type SH-SY5Y treated with CCCP cell lysates with no signal observed at this size in PINK1 knockout cell line (ab280876).

    • Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (ab300624)
      Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (ab300624)
      All lanes : Anti-PINK1 antibody [MJF-R32-7] (ab300623) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 Vehicle Control CCCP, ab141229 (0 µM, 24 h) cell lysate
      Lane 2 : Wild-type HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate
      Lane 3 : PINK1 knockout HEK-293 Vehicle Control CCCP, ab141229 (0 µM, 24 h) cell lysate
      Lane 4 : PINK1 knockout HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 63 kDa



      This data was developed using ab300623, the same antibody clone in a different buffer formulation. 

      Anti-PINK1 antibody [MJF-R32-7] (ab300623) staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. ab300623 was shown to bind specifically to PINK1. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in PINK1 knockout cell line ab266393 (knockout cell lysate ab257030). To generate this image, wild-type and PINK1 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 4 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    • Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)
      Western blot - Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)
      All lanes : Anti-PINK1 antibody [MJF-R32-7] (ab300623) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) treated with 30uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 6 hours, whole cell lysate
      Lane 2 : PINK1 knockout HEK-293T treated with 30uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 6 hours, whole cell lysate
      Lane 3 : Untreated PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : PC-3 treated with 30uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 6 hours, 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

      Performed under reducing conditions.

      Predicted band size: 63 kDa
      Observed band size: 62 kDa why is the actual band size different from the predicted?



      This data was developed using ab300623, the same antibody clone in a different buffer formulation. 

      Blocking and dilution buffer: 5% NFDM/TBST.

      In Western blot, ab300623 was shown to bind specifically to PINK1. A band was observed at 63 kDa in wild-type HEK-293T (CCCP-treated) cell lysates whereas no signal observed at this size in PINK1 knockout cell line ab266393 (CCCP-treated). 

      Exposure times: Lane 1-2: 3 minutes; Lane 3-4: 70 seconds.

    • Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)
      Anti-PINK1 antibody [MJF-R32-7] (BSA and Azide free) (AB300624)

    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 (0)

    ab300624 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。

    ab300624 は論文での使用が確認できていません。

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