参阅全部 Cathepsin B 一抗
描述兔多克隆抗体to Cathepsin B
经测试应用适用于: ICC/IF, WBmore details
- 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.
存放说明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.
存储溶液Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
纯度Immunogen affinity purified
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.
|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).|
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
- Amyloid precursor protein secretase antibody
- APP secretase antibody
- APPS antibody
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.
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.