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AB257441

Human FHL2 knockout HeLa cell lysate

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FHL2 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 deletion in exon3 and 1 bp insertion in exon3.
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Western blot - Human FHL2 knockout HeLa cell lysate (AB257441)
  • WB

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Western blot - Human FHL2 knockout HeLa cell lysate (AB257441)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : FHL2 knockout HeLa cell lysate (20 μg)

Lane 3 : K562 cell lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab202584 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.

ab202584 Recombinant Anti-FHL2 antibody [EPR17860-20] was shown to specifically react with FHL2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265475 (knockout cell lysate ab257441) was used. Wild-type and FHL2 knockout samples were subjected to SDS-PAGE. ab202584 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-FHL2 antibody [EPR17860-20] (<a href='/en-us/products/primary-antibodies/fhl2-antibody-epr17860-20-ab202584'>ab202584</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

FHL2 knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human FHL2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-fhl2-knockout-hela-cell-line-ab265475'>ab265475</a>)

Lane 3:

K562 lysate at 20 µg

Predicted band size: 32 kDa

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Western blot - Human FHL2 knockout HeLa cell lysate (AB257441)
  • WB

Lab

Western blot - Human FHL2 knockout HeLa cell lysate (AB257441)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : FHL2 knockout HeLa cell lysate (20 μg)

Lane 3 : K562 cell lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.

ab202586 Recombinant Anti-FHL2 antibody [EPR17860-23] was shown to specifically react with FHL2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265475 (knockout cell lysate ab257441) was used. Wild-type and FHL2 knockout samples were subjected to SDS-PAGE. ab202586 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-FHL2 antibody [EPR17860-23] (<a href='/en-us/products/primary-antibodies/fhl2-antibody-epr17860-23-ab202586'>ab202586</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

FHL2 knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human FHL2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-fhl2-knockout-hela-cell-line-ab265475'>ab265475</a>)

Lane 3:

K562 lysate at 20 µg

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (AB257441)
  • Sanger seq

Unknown

Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (AB257441)

Allele-1 : 1 bp insertion in exon3

Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (AB257441)
  • Sanger seq

Unknown

Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (AB257441)

Allele-2 : 1 bp deletion in exon3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon3 and 1 bp insertion in exon3.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB202584

Anti-FHL2 antibody [EPR17860-20]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-FHL2 antibody [EPR17860-20] (AB202584)

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Recombinant human VEGF 165A protein (Active)

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AB288799

Recombinant Human VCAM1 protein (Active)

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Biological Activity - Recombinant Human VCAM1 protein (Active) (AB288799)

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Cell Lines & Lysates

AB265475

Human FHL2 knockout HeLa cell line

Advanced Validation

Western blot - Human FHL2 knockout HeLa cell line (AB265475)

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  • 0
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Reactivity data

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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
FHL2
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.

The FHL2 protein also known as Four and a Half LIM Domains Protein 2 plays an important role in cellular mechanics. Its molecular mass is approximately 32 kDa. FHL2 is expressed in various tissues with significant presence in cardiac and skeletal muscles as well as the liver. It is involved in the regulation of gene expression by binding to other proteins and DNA through its LIM domains. This ability allows FHL2 to influence cellular structures and functions integrally.
Biological function summary

The FHL2 protein serves critical functions in muscle development and differentiation. It operates as part of a multiprotein complex that interacts with cytoskeletal components contributing importantly to muscle cell adhesion and communication. Moreover FHL2 engages in transcriptional regulation and acts as a coactivator or corepressor of specific transcription factors influencing gene expression patterns relevant to muscle function and maintenance.

Pathways

The FHL2 protein plays significant roles in Wnt and MAPK signaling pathways. In the Wnt signaling pathway FHL2 regulates the transcriptional activity of beta-catenin an essential component of the pathway involved in cell growth and differentiation. Additionally FHL2 contributes to the MAPK signaling pathway by influencing interactions between upstream regulators and downstream targets modulating cellular responses to external stimuli. Through these pathways FHL2 interacts with various proteins including GATA4 and TEF-1 which are vital for its regulatory functions.

FHL2 has associations with certain cancers like prostate cancer and cardiomyopathy. In prostate cancer FHL2 overexpression leads to increased tumor progression and is often linked to androgen receptors promoting malignant transformation. In cardiomyopathy alterations in FHL2 expression affect heart muscle integrity and function sometimes in conjunction with interactions involving desmin an important structural protein. Understanding FHL2's connections to these diseases offers insights into potential therapeutic targets.
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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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