Human CTSD knockout A-431 cell line
- Advanced Validation
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Human CTSD knockout A-431 cell line available to order. Recommended control: Human wild-type A-431 cell line (ab263975).
View alternative names
CATD_HUMAN, CD, CLN10, CPSD, CTSD, Cathepsin D, Cathepsin D heavy chain, Ceroid lipofuscinosis neuronal 10, Epididymis secretory sperm binding protein Li 130P, HEL S 130P, Lysosomal aspartyl peptidase, Lysosomal aspartyl protease, MGC2311
- WB
Lab
Western blot - Human CTSD knockout A-431 cell line (AB261891)
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.
All lanes:
Western blot - Anti-proCathepsin D antibody [EPR3054] (<a href='/en-us/products/primary-antibodies/procathepsin-d-antibody-epr3054-ab134169'>ab134169</a>) at 1/2000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 40 µg
Lane 2:
Cathepsin D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 40 µg
Lane 2:
Western blot - Human CTSD knockout A-431 cell line (ab261891)
Lane 3:
Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 40 µg
Lane 4:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 40 µg
False
- WB
Lab
Western blot - Human CTSD knockout A-431 cell line (AB261891)
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.
All lanes:
Western blot - Anti-Cathepsin D antibody [EPR3056Y] (<a href='/en-us/products/primary-antibodies/cathepsin-d-antibody-epr3056y-ab75811'>ab75811</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 40 µg
Lane 2:
CTSD knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 40 µg
Lane 2:
Western blot - Human CTSD knockout A-431 cell line (ab261891)
Lane 3:
Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 40 µg
Lane 4:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 40 µg
False
- WB
Lab
Western blot - Human CTSD knockout A-431 cell line (AB261891)
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.
All lanes:
Western blot - Anti-Cathepsin D antibody (<a href='/en-us/products/primary-antibodies/cathepsin-d-antibody-ab72915'>ab72915</a>) at 1 µg/mL
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
CTSD knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human CTSD knockout A-431 cell line (ab261891)
Lane 3:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4:
Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
False
- NGS
Lab
Next Generation Sequencing - Human CTSD knockout A-431 cell line (AB261891)
X = 22 bp deletion
Reactivity data
Product details
Recommended control: Human wild-type A-431 cell line (ab263975). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at Conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
Activity summary
Cathepsin D also known as CTSD is a protein with a mass of approximately 45 kDa. It functions as an aspartyl protease and is expressed in lysosomes across various tissues including the liver and kidneys. This enzyme acts by cleaving peptide bonds within proteins which is essential for protein degradation and turnover. Cathepsin D exists as precursor forms that become activated in the acidic environment of the lysosome. It plays a critical role in normal cellular processes by maintaining protein homeostasis.
Biological function summary
The enzymatic activity of Cathepsin D is important for cellular maintenance and apoptosis. This protease does not act within larger protein complexes but contributes to the degradation of extracellular and intracellular proteins. It mediates processes like antigen processing where it deconstructs proteins into peptides that are presented on major histocompatibility complex (MHC) molecules. ELISA tests can quantify its expression levels sometimes termed as CTSD activity in various biological samples offering insights into its role within cellular environments.
Pathways
Cathepsin D involvement includes the lysosomal degradation pathway and the apoptotic signaling pathway. In the lysosomal degradation pathway Cathepsin D breaks down proteins and peptides a process important for cellular recycling and energy release. It interacts with other lysosomal enzymes such as Cathepsin B in this pathway ensuring comprehensive breakdown of cellular waste. The apoptotic signaling pathway involves the regulation of programmed cell death where Cathepsin D can influence the activation of downstream proteins like Bcl-2 and Bax which control cell survival.
Associated diseases and disorders
The overexpression of Cathepsin D links to breast cancer and Alzheimer's disease. In breast cancer increased Cathepsin D expression correlates with tumor progression and metastasis influencing tumor behavior through interactions with other proteins involved in cell proliferation. Alzheimer's disease features the involvement of Cathepsin D in the breakdown of amyloid precursor protein which relates to amyloid beta plaque accumulation. The abnormal activity of Cathepsin D in these disorders makes it a potential target for therapeutic antibodies such as CTSD antibodies which aim to regulate its function.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Product protocols
- Visit the General protocols
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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