AB255377

Human CDH2 (N Cadherin) knockout HEK-293T cell line

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Human CDH2 (N Cadherin) knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).

View Alternative Names

CADH2_HUMAN, CD325, CD325 antigen, CDH2, CDHN, CDw325, CDw325 antigen, Cadherin 2 N cadherin neuronal, Cadherin 2 type 1, Cadherin 2 type 1 N cadherin neuronal, Cadherin 2, type 1, N-cadherin (neuronal), Cadherin-2, Calcium dependent adhesion protein neuronal, N cadherin 1, N-cadherin, NCAD, Neural cadherin, OTTHUMP00000066304, OTTHUMP00000067378

5 Images

Lab

Western blot - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Lanes 1 - 2 : Merged signal (red and green). Green - ab245117 observed at 125 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab245117 was shown to react with N Cadherin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255377 (knockout cell lysate ab263843) was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab245117 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-N Cadherin antibody [EPR22397-264] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr22397-264-ab245117'>ab245117</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

CDH2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CDH2 (N Cadherin) knockout HEK-293T cell line (ab255377)

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 1/20000 dilution

Predicted band size: 99 kDa

Observed band size: 125 kDa,130 kDa

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Lab

Western blot - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Lanes 1 - 2 : Merged signal (red and green). Green - ab76011 observed at 125 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab76011 was shown to react with N Cadherin in wild-type HEK-293T. Loss of signal was observed when knockout cell line ab255377 (knockout cell lysate ab263843) was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab76011 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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-N Cadherin antibody [EPR1791-4] (<a href='/en-us/products/primary-antibodies/n-cadherin-antibody-epr1791-4-ab76011'>ab76011</a>) at 1/5000 dilution

Lane 1:

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

Lane 2:

CDH2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CDH2 (N Cadherin) knockout HEK-293T cell line (ab255377)

Secondary

All lanes:

Predicted band size: 99 kDa

Observed band size: 100 kDa,125 kDa

false

Sanger Sequencing - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Sanger seq

Unknown

Sanger Sequencing - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Allele-2 : 5 bp insertion in exon 4.

Sanger seq

Unknown

Sanger Sequencing - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Allele-1 : 1 bp insertion in exon 4

Immunocytochemistry/ Immunofluorescence - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human CDH2 (N Cadherin) knockout HEK-293T cell line (AB255377)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% Tween-20 permeabilized CDH2 KO HEK-293T (CDH2 knockout human embryonic kidney epithelial cell) ab255377 cells labelling N Cadherin with ab321897 at 1/50 (9.82 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in wildtype HEK-293T cells showing no staining in CDH2 knockout HEK-293T cells(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

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, 1 bp insertion in exon 4 and 5 bp insertion in exon 4

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Reactivity data

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Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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, 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?

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Properties and storage information

Gene name

CDH2

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

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.

N-Cadherin also known as CDH2 or neuronal cadherin is a calcium-dependent cell adhesion protein. It is characterized by its role in mediating intercellular connections primarily through its presence on neurons fibroblasts and certain epithelial cells. N-Cadherin has an approximate mass of 130 kDa. The protein is integral in forming homophilic interactions where N-Cadherin molecules on adjacent cells interact to establish robust cell-cell adhesion.

Biological function summary

N-Cadherin significantly influences cell-cell interaction and communication facilitating cellular adhesion and signal transduction. It is a vital component of adherens junctions contributing to tissue morphogenesis and stability. N-Cadherin interacts with other cytoplasmic proteins such as catenins forming a complex essential for linking the actin cytoskeleton to the cell membrane. This interaction affects cellular behaviors including migration and differentiation.

Pathways

N-Cadherin plays a significant role in the neural development and epithelial-to-mesenchymal transition (EMT) pathways important for development and cancer progression. It engages with related proteins such as beta-catenin which helps transduce signals within these pathways. N-Cadherin's interactions within these pathways highlight its role in maintaining multicellular structure and signaling processes important for development and pathogenesis.

N-Cadherin is associated with cancer and heart disease. In cancer particularly in the context of the EMT N-Cadherin facilitates tumor progression and metastasis cooperating with proteins like Twist and Snail to promote these processes. In heart disease alterations in N-Cadherin expression and function can impact cardiac muscle integrity and syncytium highlighting potential interactions with connexin43 to maintain cardiac function. Understanding these associations provides insights into therapeutic targets for these conditions.

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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

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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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

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

For licensing inquiries, please contact partnerships@abcam.com