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AB263834

Human SMAD3 knockout HeLa cell lysate

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SMAD3 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 insertion in exon 4.
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Sanger Sequencing - Human SMAD3 knockout HeLa cell lysate (AB263834)
  • Sanger seq

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Sanger Sequencing - Human SMAD3 knockout HeLa cell lysate (AB263834)

Homozygous : 1 bp insertion in exon 4

Western blot - Human SMAD3 knockout HeLa cell lysate (AB263834)
  • WB

Lab

Western blot - Human SMAD3 knockout HeLa cell lysate (AB263834)

Lane 1 : Wild-type A549 cell lysate (20 μg)

Lane 2 : SMAD3 knockout A549 cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : SMAD3 knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab208182 observed at 55 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab208182 was shown to react with Smad3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255431 (knockout cell lysate ab263834) was used. Wild-type and Smad3 knockout samples were subjected to SDS-PAGE. ab208182 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-Smad3 antibody [EPR19686] - ChIP Grade (<a href='/en-us/products/primary-antibodies/smad3-antibody-epr19686-chip-grade-ab208182'>ab208182</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SMAD3 knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

Western blot - Human SMAD3 knockout HeLa cell lysate (ab263834)

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 48 kDa

Observed band size: 37 kDa,74 kDa

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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, Homozygous: 1 bp insertion in exon 4.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB40854

Anti-Smad3 antibody [EP568Y]

20ul selling size|Recombinant|KO Validated|Advanced Validation|RabMAb

Western blot - Anti-Smad3 antibody [EP568Y] (AB40854)

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Proteins & Peptides

AB85329

Recombinant Human Smad2 protein

SDS-PAGE - Recombinant Human Smad2 protein (AB85329)

  • 0
  • 0
Proteins & Peptides

AB89353

Recombinant Human Smad3 protein

Sandwich ELISA - Recombinant Human Smad3 protein (AB89353)

  • 0
  • 0
Primary Antibodies

AB191385

Anti-SIRT6 antibody [EPR18463]

RabMAb|Recombinant|KO Validated

Western blot - Anti-SIRT6 antibody [EPR18463] (AB191385)

  • 5
  • 2
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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
SMAD3
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
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Supplementary information

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

Smad3 also known as Mothers against decapentaplegic homolog 3 is a protein that plays a mechanical role in signal transduction. It acts mainly as a transcription factor and gets activated through phosphorylation. The molecular weight of Smad3 is approximately 48 kDa. It is expressed widely across numerous tissues including the cellular nucleus where it executes its function after activation.
Biological function summary

Smad3 acts as a mediator of signal transduction for the TGF-beta (transforming growth factor-beta) superfamily forming a complex with phosphorylated Smad2. This enables it to regulate transcriptional activity influencing cell proliferation differentiation and apoptosis. Smad3 also participates in various cellular processes by interacting with other co-factors and regulatory proteins that aid in fine-tuning its function.

Pathways

Smad3 plays an important role in the TGF-beta signaling pathway where it works closely with Smad4 to propagate the signal. Upon phosphorylation it forms a complex with co-Smad (Smad4) and moves into the nucleus to influence gene expression. Smad3 is also involved in pathways related to oncogenesis and tissue fibrosis indicating its significant role in cellular regulation and response mechanisms.

Smad3 is associated with fibrotic diseases and cancers particularly in tissues such as the liver and lungs. Altered Smad3 signaling contributes to the pathological process occurring in fibrotic disorders often interacting with Smad4 in these abnormalities. Dysregulated Smad3 expression or mutations can also lead to oncogenic transformations highlighting its critical involvement in disease states.
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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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