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AB263208

Human FZR1 knockout HEK-293T cell lysate

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FZR1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon9.
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Western blot - Human FZR1 knockout HEK-293T cell lysate (AB263208)
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Western blot - Human FZR1 knockout HEK-293T cell lysate (AB263208)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : FZR1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate (20 ug)

ab77885 was shown to specifically react with FZR1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab267263 (knockout cell lysate ab263208) was used. Wild-type and FZR1 knockout samples were subjected to SDS-PAGE. ab77885 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4oC at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-FZR1 antibody [AR38.2] (<a href='/en-us/products/primary-antibodies/fzr1-antibody-ar382-ab77885'>ab77885</a>) at 1/500 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

FZR1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human FZR1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-fzr1-knockout-hek-293t-cell-line-ab267263'>ab267263</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

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Sanger Sequencing - Human FZR1 knockout HEK-293T cell lysate (AB263208)
  • Sanger seq

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Sanger Sequencing - Human FZR1 knockout HEK-293T cell lysate (AB263208)

Homozygous : 1 bp insertion in exon9

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon9.

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Complementary Products

Cell Lines & Lysates

AB267263

Human FZR1 knockout HEK-293T cell line

Advanced Validation

Western blot - Human FZR1 knockout HEK-293T cell line (AB267263)

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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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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
FZR1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

FZR1 also known as fizzy-related protein homolog plays a critical mechanical role in cell cycle regulation as a co-activator of the anaphase-promoting complex/cyclosome (APC/C). The FZR1 protein has a mass of about 83 kDa. It is involved in the transition from metaphase to anaphase during cell division. This protein expresses in various tissues but shows higher levels in proliferating cells due to its role in cell cycle control.
Biological function summary

FZR1 helps regulate the degradation of cell cycle proteins by tagging them for destruction in a process involving the ubiquitin-proteasome system. FZR1 forms a complex with APC/C pushing the progression of the cell cycle and ensuring proper chromosome segregation. This activity prevents genomic instability by maintaining the correct order of cell cycle phases.

Pathways

Researchers place FZR1 in the cell cycle pathway particularly in regulating the mitotic exit and G1/S transition. One significant pathway involves its partnership with the cyclin B1 protein where FZR1 aids in its ubiquitination and subsequent degradation. It is also related to pathways involving the retinoblastoma protein (Rb) which adds a layer of regulation over the cycle and ensures DNA replication precision.

FZR1 associates with cancer due to its involvement in controlling cell division and preventing aneuploidy. Disruption in FZR1 function can lead to misregulated cell growth which is often observed in tumors. Furthermore some studies connect FZR1 to neurodegenerative disorders notably through its interactions with proteins in the nervous system although these connections are still under investigation.
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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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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

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

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