Human CTSD knockout A-431 cell lysate (ab261700)
Overview
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Product name
Human CTSD knockout A-431 cell lysate
See all Cathepsin D kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 22 bp deletion; Frameshift = 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280450 - Human CTSD knockout A431 cell lysate 1 x 100µg ab263973 - Human wild-type A-431 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
Target
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Function
Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease. -
Tissue specificity
Expressed in the aorta extrcellular space (at protein level). -
Involvement in disease
Ceroid lipofuscinosis, neuronal, 10 -
Sequence similarities
Belongs to the peptidase A1 family.
Contains 1 peptidase A1 domain. -
Post-translational
modificationsN- and O-glycosylated. -
Cellular localization
Lysosome. Melanosome. Secreted, extracellular space. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. In aortic samples, detected as an extracellular protein loosely bound to the matrix (PubMed:20551380). - Information by UniProt
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Alternative names
- CatD
- CATD_HUMAN
- Cathepsin D
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261700 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Western blot - Human CTSD knockout A-431 cell lysate (ab261700)Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 40 ug
Lane 2: Cathepsin D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 40 ug
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 40 ug
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 40 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab134169 observed at 46 kDa. Red - loading control, ab7291 (mouse anti-tubulin), observed at 50 kDa.
ab134169 was shown to recognize in wild-type A-431 cells as signal was lost at the expected MW in CTSD knockout cell line ab261891 (knockout cell lysate ab261700). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF milk. Ab134169 and ab7291 (Mouse anti-tubulin loading control) were incubated overnight at 4°C at 1/2000 dilution 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. -
Western blot - Human CTSD knockout A-431 cell lysate (ab261700)Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 40 ug
Lane 2: CTSD knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 40 ug
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 40 ug
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 40 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab75811 observed at 44 kDa. Red - loading control, ab7291, observed at 55 kDa.
ab75811 was shown to specifically react with in wild-type A-431 cells as signal was lost in CTSD knockout cell line ab261891 (knockout cell lysate ab261700). Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF milk. Ab75811 and ab7291 (Mouse anti Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution 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. -
Western blot - Human CTSD knockout A-431 cell lysate (ab261700)Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 2: CTSD knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 3: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug
Lane 4: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab72915 observed at 28, 43, 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab72915 was shown to specifically react with CTSD in wild-type A-431 cells as signal was lost in CTSD knockout cell line ab261891 (knockout cell lysate ab261700). Wild-type and CTSD knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab72915 and ab8245 (Mouse anti-GAPDH 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. -
Next Generation Sequencing - Human CTSD knockout A-431 cell lysate (ab261700)X = 22 bp deletion
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab261700 has not yet been referenced specifically in any publications.