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AB266175

Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line

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IPO4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 4 and 1 bp insertion in exon 4.

View Alternative Names

IPO4_HUMAN, Imp4, Imp4b, Importin-4, Importin-4b, Ran-binding protein 4, RanBP4

3 Images
Western blot - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)
  • WB

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Western blot - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)

Lanes 1-4 : Merged signal (red and green). Green - ab181046 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.

ab181046 Anti-Importin4/Imp4 antibody [EPR13660] was shown to specifically react with Importin4/Imp4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266175 (knockout cell lysate ab258005) was used. Wild-type and Importin4/Imp4 knockout samples were subjected to SDS-PAGE. ab181046 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-Importin4/Imp4 antibody [EPR13660] (<a href='/en-us/products/primary-antibodies/importin4-imp4-antibody-epr13660-ab181046'>ab181046</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

IPO4 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (ab266175)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

A549 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: 119 kDa

Observed band size: 130 kDa

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Sanger Sequencing - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)
  • Sanger seq

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Sanger Sequencing - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)

Allele-2 : 1 bp insertion in exon 4.

Sanger Sequencing - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)
  • Sanger seq

Unknown

Sanger Sequencing - Human IPO4 (Importin4/Imp4) knockout HEK-293T cell line (AB266175)

Allele-1 : 10 bp deletion in exon4

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, 10 bp deletion in exon 4 and 1 bp insertion in exon 4

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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, 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
IPO4
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.

Importin4 (Imp4) also known as Ipo4 is a nuclear import receptor with an approximate mass of 130 kDa. This protein facilitates the transport of molecules such as ribosomal proteins and histones into the nucleus. Expression of Importin4 occurs in various tissues including the brain testis and liver. Importin4 plays a significant mechanical role by recognizing nuclear localization signals on cargo molecules and mediating their translocation across the nuclear envelope.
Biological function summary

Importin4 functions as part of the nuclear transport system a complex network essential for cellular operations. Importin4 interacts with the nuclear pore complex to enable import of specific protein cargo into the nucleus. By partnering with karyopherins and Ran-GTPase Importin4 participates in the active transport mechanism ensuring the correct localization of proteins that are critical to genomic regulation processes.

Pathways

Importin4 plays a role in the nuclear transport pathway and is associated with the ribosome biogenesis pathway. This protein works alongside proteins like Ran-GTPase enhancing the translocation of ribosomal components pivotal for cellular growth and protein synthesis. Importin4 also aids in the shuttling of histones which are essential to chromatin assembly and DNA transcription regulation.

Importin4 has connections to cancer and neurodegenerative diseases. Altered expression and mutations in Importin4 may disrupt cellular homeostasis and nuclear transport linking it to tumorigenesis. Additionally associations with proteins such as karyopherin alpha importers indicate its involvement in pathological states affecting the nervous system potentially contributing to conditions such as Alzheimer's disease. Understanding Importin4's interactions could provide insights into these complex diseases.
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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 IPO4
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Product promise

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