Anti-Cathepsin B antibody (ab92955)
Key features and details
- Rabbit polyclonal to Cathepsin B
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Cathepsin B antibody
See all Cathepsin B primary antibodies -
Description
Rabbit polyclonal to Cathepsin B -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human Cathepsin B aa 200-300.
(Peptide available asab103276) -
Positive control
- 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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications OrangutanApplication Abreviews Notes ICC/IF Use a concentration of 1 µg/ml.WB (1) Use a concentration of 1 µg/ml. Detects a band of approximately 30, 25 kDa (predicted molecular weight: 38 kDa).Notes 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).Target
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Function
Thiol protease which is believed to participate in intracellular degradation and turnover of proteins. Has also been implicated in tumor invasion and metastasis. -
Sequence similarities
Belongs to the peptidase C1 family. -
Cellular localization
Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 1508 Human
- Omim: 116810 Human
- SwissProt: P07858 Human
- Unigene: 520898 Human
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Alternative names
- Amyloid precursor protein secretase antibody
- APP secretase antibody
- APPS antibody
see all
Images
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Western blot - Anti-Cathepsin B antibody (ab92955)
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)
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Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin B antibody (ab92955)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.
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Western blot - Anti-Cathepsin B antibody (ab92955)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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab92955 has been referenced in 3 publications.
- Blauwendraat C et al. Genetic modifiers of risk and age at onset in GBA associated Parkinson's disease and Lewy body dementia. Brain 143:234-248 (2020). PubMed: 31755958
- Martínez-Fábregas J et al. Lysosomal protease deficiency or substrate overload induces an oxidative-stress mediated STAT3-dependent pathway of lysosomal homeostasis. Nat Commun 9:5343 (2018). PubMed: 30559339
- 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). PubMed: 27932519
Images
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Western blot - Anti-Cathepsin B antibody (ab92955)
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)
-
Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin B antibody (ab92955)
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.
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Western blot - Anti-Cathepsin B antibody (ab92955)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.
