概述

  • 产品名称
    Anti-Cathepsin B抗体
    参阅全部 Cathepsin B 一抗
  • 描述
    兔多克隆抗体to Cathepsin B
  • 宿主
    Rabbit
  • 经测试应用
    适用于: ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Orangutan
  • 免疫原

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

    参阅Abcam的专有抗源政策 Synthetic peptide of Human Cathepsin B.
    参阅Abcam的专有抗源政策

  • 阳性对照
    • 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.

性能

应用

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.

应用 Ab评论 说明
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).

靶标

  • 功能
    Thiol protease which is believed to participate in intracellular degradation and turnover of proteins. Has also been implicated in tumor invasion and metastasis.
  • 序列相似性
    Belongs to the peptidase C1 family.
  • 细胞定位
    Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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 (Negative Control) (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
    All lanes : 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 seconds


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

文献

This product has been referenced in:
  • Pinheiro PV  et al. Host Plants Indirectly Influence Plant Virus Transmission by Altering Gut Cysteine Protease Activity of Aphid Vectors. Mol Cell Proteomics 16:S230-S243 (2017). Read more (PubMed: 27932519) »
See 1 Publication for this product

客户评价及客户问答

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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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提交于 Nov 11 2016

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