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AB255368

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

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CANX KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (AB255368)
  • WB

Lab

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

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 (ab255368)

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

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

Lab

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

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 37 kDa.

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 µg/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 (ab255368)

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

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

Lab

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

Lanes 1- 2 : Merged signal (red and green). Green - ab133615 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab133615 was shown to 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 HEK-293T and CANX knockout HEK-293T cell lysates 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) overnight at 4°C at a 1 in 1000 dilution and a 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 (ab255368)

Predicted band size: 68 kDa

Observed band size: 90 kDa

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Sanger Sequencing - Human CANX (Calnexin) knockout HEK-293T cell line (AB255368)
  • Sanger seq

Unknown

Sanger Sequencing - Human CANX (Calnexin) knockout HEK-293T cell line (AB255368)

Homozygous : 19 bp deletion in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 2

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Complementary Products

Primary Antibodies

AB225062

Alexa Fluor® 647 Anti-Calnexin antibody [EPR3632]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Calnexin antibody [EPR3632] (AB225062)

  • 0
  • 0
Primary Antibodies

AB125011

Anti-TSG101 antibody [EPR7130(B)]

BOND RX™ Validated|RabMAb|Recombinant

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

  • 4
  • 10
Cell Lines & Lysates

AB263805

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

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

  • 0
  • 0
Primary Antibodies

AB134045

Anti-CD63 antibody [EPR5702] - Late Endosome Marker

RabMAb|Recombinant|KO Validated|Lab Essentials|Trial Size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] - Late Endosome Marker (AB134045)

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  • 6
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Reactivity data

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Product details

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.

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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
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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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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Target data

See full target information CANX
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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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