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AB257411

Human DIAPH1 knockout HEK-293T cell lysate

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DIAPH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

View Alternative Names

DFNA1, DIA1, DIAP1_HUMAN, DRF1, Diaphanous homolog 1 (Drosophila), Diaphanous-related formin-1, FLJ25265, LFHL1, Protein diaphanous homolog 1, deafness, autosomal dominant 1, diaphanous homolog 1, hDIA1, low frequency hearing loss 1, p140DIA

2 Images
Western blot - Human DIAPH1 knockout HEK-293T cell lysate (AB257411)
  • WB

Lab

Western blot - Human DIAPH1 knockout HEK-293T cell lysate (AB257411)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : DIAPH1 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab129167 observed at 150 kDa. Red - loading control ab7291 observed at 50 kDa.

ab129167 Anti-DIAPH1 antibody [EPR7948] was shown to specifically react with DIAPH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266120 (knockout cell lysate ab257411) was used. Wild-type and DIAPH1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab129167 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 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-DIAPH1 antibody [EPR7948] (<a href='/en-us/products/primary-antibodies/diaph1-antibody-epr7948-ab129167'>ab129167</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

DIAPH1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human DIAPH1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-diaph1-knockout-hct116-cell-line-ab273727'>ab273727</a>)

Predicted band size: 141 kDa

Observed band size: 150 kDa

false

Sanger Sequencing - Human DIAPH1 knockout HEK-293T cell lysate (AB257411)
  • Sanger seq

Unknown

Sanger Sequencing - Human DIAPH1 knockout HEK-293T cell lysate (AB257411)

Homozygous : 1 bp deletion in exon 1

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, Homozygous: 1 bp deletion in exon 1.

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 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.

What's included?

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Properties and storage information

Gene name
DIAPH1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

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

DIAPH1 also known as Diaphanous-related formin 1 operates mainly in actin polymerization and is critical for filament formation. It weighs approximately 140 kDa. DIAPH1 expresses in a variety of tissues including heart liver and kidney making it a target of interest in multiple biological contexts. The protein plays a role in maintaining proper cytoskeletal dynamics by regulating the assembly of actin filaments.
Biological function summary

DIAPH1 functions as part of the formin family which are important in cytoskeletal organization. It interacts with other proteins to initiate and elongate actin filaments often acting within larger protein complexes that control cellular morphology and movement. DIAPH1 contributes to muscle contraction cell division and cell migration highlighting its importance in cellular structure and dynamics.

Pathways

DIAPH1 plays a significant role in the Rho GTPase signaling pathway. This pathway connects to cytoskeleton remodeling impacting cell shape and movement. DIAPH1 associates with proteins like RhoA which regulates its activity. Additionally DIAPH1 influences the Wnt signaling pathway known for its involvement in cell proliferation and differentiation.

DIAPH1 is connected to a form of sensorineural hearing loss and leukemia. Mutations in DIAPH1 can lead to autosomal dominant deafness as it affects the proper maintenance of actin structures in hair cells within the inner ear. The protein also interacts with genes linked to cancer development including those coding for p53 influencing tumorigenesis and cell cycle regulation.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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

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