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AB275252

Human DIAPH1 knockout HCT116 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: 2 bp deletion in exon 1.
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Western blot - Human DIAPH1 knockout HCT116 cell lysate (AB275252)
  • WB

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Western blot - Human DIAPH1 knockout HCT116 cell lysate (AB275252)

Lane 1 : Wild-type HCT116 cell lysate 20 ug
Lane 2 : DIAPH1 knockout HCT116 cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab129167 observed at 155 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab129167 was shown to react with DIAPH1 in wild-type HCT 116 cells in western blot with loss of signal observed in DIAPH1 knockout cell line ab273727 (DIAPH1 knockout cell lysate ab275252). HCT 116 wild-type and DIAPH1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab129167 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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 HCT116 cell lysate at 20 µg

Lane 2:

DIAPH1 knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human DIAPH1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-diaph1-knockout-hek-293t-cell-line-ab266120'>ab266120</a>)

Predicted band size: 141 kDa

Observed band size: 155 kDa

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Sanger Sequencing - Human DIAPH1 knockout HCT116 cell lysate (AB275252)
  • Sanger seq

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Sanger Sequencing - Human DIAPH1 knockout HCT116 cell lysate (AB275252)

Allele-1 : 2 bp deletion in exon 1 .

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 1

Disease

Carcinoma

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Complementary Products

Cell Lines & Lysates

AB275249

Human TRIB3 knockout HCT116 cell lysate

Western blot - Human TRIB3 knockout HCT116 cell lysate (AB275249)

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Primary Antibodies

AB129167

Anti-DIAPH1 antibody [EPR7948]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-DIAPH1 antibody [EPR7948] (AB129167)

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  • 1
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

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Proteins & Peptides

AB280753

Recombinant Human PD1 protein

Mass Spectrometry - Recombinant Human PD1 protein (AB280753)

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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 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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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
Dry 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.

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.
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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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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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