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AB263805

Human CANX (Calnexin) knockout HEK-293T cell lysate

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CANX KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon2.
4 Images
Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)
  • WB

Lab

Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)

Lane 1 : Wild-type HEK-293T cell lysate (20 ug)
Lane 2 : CANX knockout HEK-293T cell lysate (20 ug)
Lane 3 : U-2 OS cell lysate (20 ug)
Lane 4 : MCF7 cell lysate (20 ug)

Lanes 1 - 4 : Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab92573 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92573 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 20000 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-Calnexin antibody [EPR3632] (<a href='/en-us/products/primary-antibodies/calnexin-antibody-epr3632-ab92573'>ab92573</a>) at 1/20000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

CANX knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-canx-calnexin-knockout-hek-293t-cell-line-ab255368'>ab255368</a>)

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 68 kDa

Observed band size: 80 kDa

false

Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)
  • WB

Lab

Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)

Lane 1 : wild-type HEK-293T cell lysate 20 ug
Lane 2 : CANX knockout HEK-293T cell lysate 20 ug
Lane 3 : U-2 OS cell lysate 20 ug
Lane 4 : MCF7 cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab238078 observed at 80 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab238078 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab238078 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 1 ug/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Calnexin antibody [CANX/1543] (<a href='/en-us/products/primary-antibodies/calnexin-antibody-canx-1543-ab238078'>ab238078</a>) at 1 µg/mL

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

CANX knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-canx-calnexin-knockout-hek-293t-cell-line-ab255368'>ab255368</a>)

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 68 kDa

Observed band size: 80 kDa

false

Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)
  • WB

Lab

Western blot - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : CANX knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab133615 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133615 Recombinant Anti-Calnexin antibody [EPR3632] was shown to specifically react with CANX in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255368 (knockout cell lysate ab263805) was used. Wild-type and CANX knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133615 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (<a href='/en-us/products/primary-antibodies/calnexin-antibody-epr36332-er-membrane-marker-ab133615'>ab133615</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

CANX knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-canx-calnexin-knockout-hek-293t-cell-line-ab255368'>ab255368</a>)

Predicted band size: 68 kDa

Observed band size: 90 kDa

false

Sanger Sequencing - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)
  • Sanger seq

Unknown

Sanger Sequencing - Human CANX (Calnexin) knockout HEK-293T cell lysate (AB263805)

Homozygous : 19 bp deletion in exon2

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, 19 bp deletion in exon2.

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Complementary Products

Cell Lines & Lysates

AB255368

Human CANX (Calnexin) knockout HEK-293T cell line

Advanced Validation

Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (AB255368)

  • 0
  • 0
Primary Antibodies

AB125011

Anti-TSG101 antibody [EPR7130(B)]

BOND RX™ Validated|RabMAb|Recombinant

Immunohistochemistry - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

  • 4
  • 10
Primary Antibodies

AB22595

Anti-Calnexin antibody - ER Marker

Lab Essentials|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody - ER Marker (AB22595)

  • 5
  • 28
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
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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?

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Properties and storage information

Gene name
CANX
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.

Calnexin also known as Canx is a type I integral membrane protein of the endoplasmic reticulum (ER) involved in the process of protein folding. This chaperone protein has an approximate molecular weight of 90 kDa and is known for its role in the quality control of glycoproteins. Calnexin is expressed in the ER of cells where it interacts with nascent polypeptides to ensure proper folding and assembly contributing to cellular homeostasis. It exhibits its function through its lectin-like domain that binds to sugar moieties on glycoproteins.
Biological function summary

Calnexin facilitates the proper folding of newly synthesized proteins by forming a complex with another chaperone protein called ERp57. This interaction helps in creating the correct disulfide bonds in glycoproteins which is essential for their stability and functionality. The complex often referred to as the calnexin cycle is critical in preventing the aggregation and misfolding of proteins within the ER. This process ensures that only correctly folded proteins proceed to the Golgi apparatus for further processing and transport.

Pathways

Calnexin plays an important role in the ER-associated degradation (ERAD) pathway and the unfolded protein response (UPR). In these pathways calnexin ensures that misfolded proteins are retained in the ER or targeted for degradation preventing cellular stress. Calnexin is associated with proteins such as calreticulin another chaperone protein with a similar function in the ER. Together they maintain proteostasis within cells and protect against the accumulation of improperly folded proteins.

Calnexin is linked to several conditions including cystic fibrosis and certain neurodegenerative diseases. In cystic fibrosis the misfolding and subsequent degradation of the CFTR protein are associated with calnexin's role in the ERAD pathway. Similarly in neurodegenerative diseases such as Alzheimer's disrupted protein folding and aggregation are linked to ER stress where calnexin and other chaperone proteins like BiP play a pivotal role in managing protein misfolding. Understanding calnexin's role in these disorders can contribute to developing strategies to mitigate faulty protein folding and its pathological consequences.
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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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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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