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AB266428

Human TRIP10 (Cip4) knockout HEK-293T cell line

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TRIP10 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 2.

View Alternative Names

CG11341, CG15015 PA, CIP4_HUMAN, Cdc42 interaction protein 4 long isoform, Cdc42-interacting protein 4, DCIP4, Protein Felic, STOT, Salt tolerant protein, Salt tolerator, TR-interacting protein 10, TRIP-10, Thyroid hormone receptor interactor 10, Thyroid receptor-interacting protein 10, Truncated Cdc42 interaction protein 4, hSTP

3 Images
Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)
  • WB

Lab

Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)

Lanes 1-4 : Merged signal (red and green). Green - ab108313 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108313 Anti-Cip4 antibody [EPR1965] was shown to specifically react with Cip4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266428 (knockout cell lysate ab258251) was used. Wild-type and Cip4 knockout samples were subjected to SDS-PAGE. ab108313 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-Cip4 antibody [EPR1965] (<a href='/en-us/products/primary-antibodies/cip4-antibody-epr1965-ab108313'>ab108313</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

TRIP10 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (ab266428)

Lane 3:

JAR cell lysate at 20 µg

Lane 4:

HepG2 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: 11 kDa,21 kDa,22 kDa,33 kDa,37 kDa,40 kDa,42 kDa,48 kDa,57 kDa,61 kDa,65 kDa,67 kDa,68 kDa,69 kDa

Observed band size: 100-180 kDa,15 kDa,21 kDa,26 kDa,33 kDa,37 kDa,40 kDa,48 kDa,49 kDa,65 kDa,67 kDa,70 kDa,75 kDa

false

Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)
  • WB

Lab

Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)

Lanes 1-4 : Merged signal (red and green). Green - ab108277 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108277 Anti-Cip4 antibody [EPR1966(2)] was shown to specifically react with Cip4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266428 (knockout cell lysate ab258251) was used. Wild-type and Cip4 knockout samples were subjected to SDS-PAGE. ab108277 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-Cip4 antibody [EPR1966(2)] (<a href='/en-us/products/primary-antibodies/cip4-antibody-epr19662-ab108277'>ab108277</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

TRIP10 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human TRIP10 (Cip4) knockout HEK-293T cell line (ab266428)

Lane 3:

JAR cell lysate at 20 µg

Lane 4:

HepG2 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: 68 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRIP10 (Cip4) knockout HEK-293T cell line (AB266428)

Homozygous : 14 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: 14 bp deletion in exon 2

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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 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
TRIP10
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.

Cip4 also known as Cdc42-interacting protein 4 is a member of the F-BAR family and is involved in cellular processes related to actin dynamics. It has a molecular mass of approximately 70 kDa. Cip4 interacts with Cdc42 a small GTPase to facilitate membrane bending and invagination important for endocytosis and membrane trafficking. This protein is expressed in a variety of tissues including the brain heart and liver reflecting its broad functional significance across different cell types.
Biological function summary

Cip4 plays an essential role in linking membranes to the actin cytoskeleton. It is part of a complex with other proteins like Toca-1 and N-WASP which are important for actin polymerization. This function supports cellular morphogenesis and migration by organizing the actin filaments in response to external signals. Cip4 also contributes to the regulation of cell shape and motility aiding in processes such as development and immune responses by modulating membrane-associated actin assembly.

Pathways

Cip4 participates in pathways including the Cdc42 signaling pathway and endocytosis. In the Cdc42 signaling pathway Cip4 works with proteins like N-WASP and dynamin to facilitate membrane remodeling and cytoskeletal rearrangements impacting processes like cell division and signal transduction. Its involvement in endocytosis influences cargo uptake and intracellular transport linking it to dynamin-dependent endocytic pathways critical for nutrient uptake and receptor internalization.

Cip4 is associated with certain cancers and neurological disorders. Its dysregulation can contribute to tumor progression often involving interactions with proteins like Cdc42 and N-WASP that promote changes in cell adhesion and motility. Additionally abnormalities in Cip4 expression or function may lead to disrupted cell signaling in neurological disorders impacting processes essential for brain development and function. Understanding Cip4's role in these contexts provides insights into potential therapeutic targets for treatment.
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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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Primary Antibodies

AB108313

Anti-Cip4 antibody [EPR1965]

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