製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Cathepsin B
  • アプリケーション
    適用あり: ICC/IF, WBmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Orangutan
  • 免疫原

    Synthetic peptide derived from within residues 200 - 300 of Human Cathepsin B.

    Synthetic peptide of Human Cathepsin B.

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following lysates: Human Liver Tissue; Human Kidney Tissue; Human Brain Tissue; Human Placenta Tissue; HepG2 Whole Cell; HEK293 Whole Cell Lysate.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab92955 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 30, 25 kDa (predicted molecular weight: 38 kDa).

ターゲット情報

  • 機能
    Thiol protease which is believed to participate in intracellular degradation and turnover of proteins. Has also been implicated in tumor invasion and metastasis.
  • 配列類似性
    Belongs to the peptidase C1 family.
  • 細胞内局在
    Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Amyloid precursor protein secretase antibody
    • APP secretase antibody
    • APPS antibody
    • CATB_HUMAN antibody
    • Cathepsin B heavy chain antibody
    • Cathepsin B1 antibody
    • CathepsinB antibody
    • CPSB antibody
    • CTSB antibody
    • cysteine protease antibody
    • OTTHUMP00000116009 antibody
    • OTTHUMP00000229510 antibody
    • OTTHUMP00000229511 antibody
    • OTTHUMP00000229512 antibody
    • OTTHUMP00000229514 antibody
    • OTTHUMP00000229515 antibody
    • OTTHUMP00000229516 antibody
    • Preprocathepsin B antibody
    see all

画像



  • Predicted band size : 38 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Cathepsin B knockout HAP1 whole cell lysate (20 µg)
    Lane 3: A549 whole cell lysate (20 µg)
    Lane 4: HL60 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab92955 observed at 24-27-44 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab92955 was shown to recognize Cathepsin B in wild-type HAP1 cells as signal was lost at the expected MW in Cathepsin B knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Cathepsin B knockout samples were subjected to SDS-PAGE. Ab92955 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml 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.

    Pro-cathepsin B runs at approximately 44 kDa on SDS-PAGE. When it is proteolytically processed and glycosylated it forms a mature two-chain protein containing a heavy chain (running at 27 and 24 kDa) and a light chain (5 kDa)

  • ICC/IF image of ab92955 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab92955 at 1µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Cathepsin B antibody (ab92955) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human kidney tissue lysate - total protein (ab30203)
    Lane 3 : Human brain tissue lysate - total protein (ab29466)
    Lane 4 : Human placenta tissue lysate - total protein (ab29745)
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 25,30 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 secondsWe hypothesize that the 25 and 30 kDa bands represent the heavy chain and mature form of Cathepsin B, respectively. Abcam welcomes customer feedback.

参考文献

ab92955 has not yet been referenced specifically in any publications.

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (283T cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
283T cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Nov 11 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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