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AB265247

Human FERMT2 (Kindlin 2) knockout HeLa cell line

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FERMT2 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: 1 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

FERM2_HUMAN, FLJ34213, FLJ44462, Fermitin family homolog 2, Fermt2, KIND2, Kindlin-2, MIG-2, Mitogen-inducible gene 2 protein, PH domain-containing family C member 1, PLEKHC1, Pleckstrin homology domain containing family C (with FERM domain) member 1, Pleckstrin homology domain-containing family C member 1, UNC112, UNC112B

2 Images
Western blot - Human FERMT2 (Kindlin 2) knockout HeLa cell line (AB265247)
  • WB

Lab

Western blot - Human FERMT2 (Kindlin 2) knockout HeLa cell line (AB265247)

Lanes 1-3 : Merged signal (red and green). Green - ab74030 observed at 78 kDa. Red - loading control ab8245 observed at 36 kDa.

ab74030 Anti-Kindlin 2 antibody was shown to specifically react with Kindlin 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265247 (knockout cell lysate ab257436) was used. Wild-type and Kindlin 2 knockout samples were subjected to SDS-PAGE. ab74030 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 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-Kindlin 2 antibody (<a href='/en-us/products/primary-antibodies/kindlin-2-antibody-ab74030'>ab74030</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FERMT2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FERMT2 (Kindlin 2) knockout HeLa cell line (ab265247)

Lane 3:

Saos-2 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: 78 kDa

Observed band size: 78 kDa

false

Sanger Sequencing - Human FERMT2 (Kindlin 2) knockout HeLa cell line (AB265247)
  • Sanger seq

Unknown

Sanger Sequencing - Human FERMT2 (Kindlin 2) knockout HeLa cell line (AB265247)

Homozygous : 1 bp deletion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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.

What's included?

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Properties and storage information

Gene name
FERMT2
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

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.

Supplementary information

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

Kindlin-2 also known as FERMT2 is a mechanistic component involved in cellular adhesion processes. This protein has a molecular weight of approximately 75 kDa. Kindlin-2 is found in various tissues with high expression in epithelial cells and muscle tissues. It works together with integrins to maintain cell adhesion to the extracellular matrix performing an important role in cell communication with its surroundings.
Biological function summary

Kindlin-2 contributes to integrin activation and cytoskeletal organization by forming part of a larger complex. This complex stabilizes integrin-talin interactions enhancing the affinity of integrins for their ligands. This function of Kindlin-2 aids in dynamically regulating cell shape migration and mechanical signaling. Its role is significant for normal cellular function and homeostasis because it helps cells to attach and detach when necessary which is important for cell movement and tissue repair.

Pathways

The action of Kindlin-2 integrates with key signaling pathways like the Wnt and PI3K/AKT pathways. In these pathways Kindlin-2 assists in transmitting signals necessary for cellular proliferation and survival. It interacts with other proteins such as talin and integrin β subunits forming a bridge between the cytoskeleton and external signals. This interaction allows cells to adapt to changes in their environment by modulating cell adhesion and migration.

Kindlin-2's function relates to conditions such as cancer and fibrosis. In cancer Kindlin-2 aids tumor cell migration and metastasis linking it to the pathological spread of cancerous cells. In fibrosis Kindlin-2 influences the excessive deposition of extracellular matrix components contributing to tissue scarring. The protein also interacts with fibronectin and collagens in these contexts showing its involvement in extracellular matrix remodeling and pathological tissue changes.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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