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AB289247

Human IGF2BP3 knockout HCT116 cell line

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IGF2BP3 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. 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

CT98, Cancer/testis antigen 98, DKFZp686F1078, IF2B3_HUMAN, IGF-II mRNA-binding protein 3, IGF2 mRNA-binding protein 3, IGF2BP3, Insulin-like growth factor 2 mRNA-binding protein 3, KH domain-containing protein overexpressed in cancer, KOC 1, VICKZ 3, VICKZ family member 3, hKOC

3 Images
Western blot - Human IGF2BP3 knockout HCT116 cell line (AB289247)
  • WB

Lab

Western blot - Human IGF2BP3 knockout HCT116 cell line (AB289247)

Western blot : Anti-IGF2BP3 antibody [EPR12022] (ab177942) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab177942 was shown to bind specifically to IGF2BP3. A band was observed at 60-75 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in IGF2BP3 knockout cell line. To generate this image, wild-type and IGF2BP3 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-IMP3 antibody [EPR12022] (<a href='/en-us/products/primary-antibodies/imp3-antibody-epr12022-ab177942'>ab177942</a>) at 1/2000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

IGF2BP3 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

IGF2BP3 knockout HAP1 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

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Next Generation Sequencing - Human IGF2BP3 knockout HCT116 cell line (AB289247)
  • NGS

Supplier Data

Next Generation Sequencing - Human IGF2BP3 knockout HCT116 cell line (AB289247)

160 bp deletion after Leu164 of the WT protein.

Sanger Sequencing - Human IGF2BP3 knockout HCT116 cell line (AB289247)
  • Sanger seq

Project

Sanger Sequencing - Human IGF2BP3 knockout HCT116 cell line (AB289247)

160bp after Leu165 in WT protein.

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Sanger Sequencing,Western blot

Disease

Carcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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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Properties and storage information

Gene name
IGF2BP3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, 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

McCoY5a + 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.

IMP3 also known as IGF2BP3 is an oncofetal protein belonging to the insulin-like growth factor 2 mRNA-binding protein family. It has a molecular weight of around 65 kDa. IMP3 expresses in various tissues including embryonic tissues and several cancer types. In healthy adults the expression is minimal observed mostly in the testis placenta and some parts of the nervous system. It's significance in development and cancer makes it an important target for research.
Biological function summary

IMP3 plays a role in the regulation of mRNA stability localization and translation. It is a part of ribonucleoprotein complexes that facilitate these processes by binding to the 3' untranslated regions of target mRNAs. The protein influences the expression of genes involved in cell growth differentiation and migration. IMP3's ability to modulate mRNA fate contributes to cellular events essential for development and has implications in cancer progression.

Pathways

The involvement of IMP3 is significant in pathways like the insulin-like growth factor (IGF) signaling pathway and epithelial-mesenchymal transition (EMT). Within these pathways IMP3 modulates the activities of proteins such as IGF2 influencing cell proliferation and survival. Its participation in EMT is essential for processes like metastasis in cancer cells linking it to broader biological processes that govern cell movement and identity change.

High IMP3 expression associates with various cancers including pancreatic and ovarian cancers. It has been connected to the progression and poor prognosis of these cancers. In pancreatic cancer IMP3 can interact with proteins like IGF2 enhancing tumor growth and metastasis. This connection makes IMP3 a potential biomarker for cancer diagnostics and therapeutic targeting.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information IGF2BP3
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