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AB261700

Human CTSD knockout A-431 cell lysate

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CTSD KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 22 bp deletion Frameshift = 100%.
4 Images
Western blot - Human CTSD knockout A-431 cell lysate (AB261700)
  • WB

Lab

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.

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 (<a href='/en-us/products/cell-lines/human-ctsd-knockout-a-431-cell-line-ab261891'>ab261891</a>)

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

Predicted band size: 44 kDa

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Western blot - Human CTSD knockout A-431 cell lysate (AB261700)
  • WB

Lab

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.

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 (<a href='/en-us/products/cell-lines/human-ctsd-knockout-a-431-cell-line-ab261891'>ab261891</a>)

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

Predicted band size: 44 kDa

Observed band size: 44 kDa

false

Western blot - Human CTSD knockout A-431 cell lysate (AB261700)
  • WB

Lab

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.

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 (<a href='/en-us/products/cell-lines/human-ctsd-knockout-a-431-cell-line-ab261891'>ab261891</a>)

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

Predicted band size: 44 kDa,45 kDa

Observed band size: 46 kDa

false

Next Generation Sequencing - Human CTSD knockout A-431 cell lysate (AB261700)
  • NGS

Lab

Next Generation Sequencing - Human CTSD knockout A-431 cell lysate (AB261700)

X = 22 bp deletion

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 22 bp deletion Frameshift = 100%

Disease

Epidermoid Carcinoma

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Assay Kits

AB65302

Cathepsin D Activity Assay Kit (Fluorometric)

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302)

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Primary Antibodies

AB75852

Anti-Cathepsin D antibody [EPR3057Y]

BOND RX™ Validated|RabMAb|Recombinant|20ul selling size

Western blot - Anti-Cathepsin D antibody [EPR3057Y] (AB75852)

  • 5
  • 11
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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. 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.

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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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab261700-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human CTSD knockout A431 cell lysate", "number":"AB261700-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A-431 cell lysate", "number":"AB261700-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
CTSD
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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