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AB257379

Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate

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CAPN2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 4 and 1 bp insertion in exon 4 and 47 bp deletion in exon 4 and 8 bp deletion in exon 4.

View Alternative Names

CALP80, CAN2_HUMAN, CANP 2, CANP L2, CANPml, CAPN 2, Calcium activated neutral proteinase, Calcium-activated neutral protease 2, catalytic subunit, Calcium-activated neutral proteinase 2, Calpain 2 (m/II) large subunit, Calpain 2 large catalytic subunit, Calpain 2, large [catalytic] subunit, Calpain M-type, Calpain large polypeptide L2, Calpain-2 catalytic subunit, Calpain-2 large subunit, Calpain2, Capa2, FLJ39928, M calpin, M type, M-calpain, Millimolar-calpain, mCANP

6 Images
Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • WB

Lab

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Lane 1 : Wild-type HEK-293T cell lysate 40 ug
Lane 2 : CAPN2 knockout HEK-293T cell lysate 40 ug
Lane 3 : A431 cell lysate 20 ug
Lane 4 : LNCaP cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab75994 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab75994 was shown to react with Calpain 2 in wild-type HEK-293T cells in Western blot with loss of signal observed in CAPN2 knockout cell line ab266628 (CAPN2 knockout cell lysate ab257379). Wild-type HEK-293T and CAPN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab75994 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR2562Y] (<a href='/en-us/products/primary-antibodies/calpain-2-antibody-epr2562y-ab75994'>ab75994</a>) at 1/2000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 40 µg

Lane 2:

CAPN2 knockout HEK-293T cell lysate at 40 µg

Lane 2:

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-capn2-calpain-2-knockout-hek-293t-cell-line-ab266628'>ab266628</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

LNCaP cell lysate at 20 µg

Predicted band size: 80 kDa

Observed band size: 75 kDa

false

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • WB

Lab

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : CAPN2 knockout HEK293T cell lysate (20 ug)
Lane 3 : A431 cell lysate (20 ug)
Lane 4 : LNCaP cell lysate (20 ug)

ab126600 was shown to specifically react with Calpain 2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266628 (knockout cell lysate ab257379) was used. Wild-type and Calpain 2 knockout samples were subjected to SDS-PAGE. ab126600 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (<a href='/en-us/products/primary-antibodies/calpain-2-antibody-epr5977-ab126600'>ab126600</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CAPN2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-capn2-calpain-2-knockout-hek-293t-cell-line-ab266628'>ab266628</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

LNCaP cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa

false

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Allele-4 : 1 bp insertion in exon 4

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Allele-2 : 17 bp deletion in exon 4

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Allele-3 : 8 bp deletion in exon 4

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAPN2 (Calpain 2) knockout HEK-293T cell lysate (AB257379)

Allele-1 : 47 bp deletion in exon 4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 4 and 1 bp insertion in exon 4 and 47 bp deletion in exon 4 and 8 bp deletion in exon 4.

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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257379-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human CAPN2 knockout HEK293T cell lysate", "number":"AB257379-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257379-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
CAPN2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger 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.

Calpain 2 also known as CAPN2 or calpain-2 is a heterodimer protease enzyme weighing around 80 kDa. It belongs to the calpain family which requires calcium ions for activation. Calpain 2 expresses in many tissues including the brain muscles and liver indicating its widespread role in cellular processes. The enzyme comprises a large catalytic subunit and a small regulatory subunit each encoded by different genes.
Biological function summary

Calpain-2 plays a significant role in cell signaling cytoskeletal remodeling and apoptosis regulation. It does not typically function in isolation but partners with other molecules forming complexes necessary for precise proteolytic activity. This enzyme modulates several cell functions by cleaving substrates like cytoskeletal proteins transcription factors and enzymes which allows cells to respond adaptively to various stimuli.

Pathways

Calpain-2 integrates into important signaling cascades such as the ERK and PI3K/AKT pathways. It influences these pathways by processing specific proteins and modifying their activity thereby affecting cell survival and proliferation. Calpain-2 closely interacts with proteins like phosphatidylinositol-45-bisphosphate 3-kinase (PI3K) and extracellular signal-regulated kinases (ERKs) within these pathways emphasizing its importance in finely-tuning cellular responses.

Calpain-2 links to several pathological conditions including neurodegenerative diseases like Alzheimer's and muscular disorders such as Duchenne Muscular Dystrophy. In Alzheimer's disease calpain-2 activity influences the processing of amyloid precursor proteins contributing to plaque formation. In Duchenne Muscular Dystrophy calpain-2 activity disrupts muscle fiber integrity through its interaction with proteins like dystrophin aggravating the disorder's progression.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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