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AB264513

Human SMAD3 knockout A549 cell lysate

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Human SMAD3 knockout A549 cell lysate suitable for WB. View our extensive range of validated lysates from normal and diseased human, mouse and rat tissue.

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DKFZP586N0721, DKFZp686J10186, HSPC193, HST17436, JV15-2, LDS1C, LDS3, MAD (mothers against decapentaplegic Drosophila) homolog 3, MAD homolog 3, MAD, mothers against decapentaplegic homolog 3, MADH 3, MGC60396, Mad homolog JV15 2, Mad protein homolog, Mothers against DPP homolog 3, Mothers against decapentaplegic homolog 3, SMA and MAD related protein 3, SMAD, SMAD family member 3, SMAD, mothers against DPP homolog 3, SMAD3_HUMAN, hMAD-3, hSMAD3

4 Images
Next Generation Sequencing - Human SMAD3 knockout A549 cell lysate (AB264513)
  • NGS

Supplier Data

Next Generation Sequencing - Human SMAD3 knockout A549 cell lysate (AB264513)

Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 100%

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2 : SMAD3 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4 : Human Kidney cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab208182 observed at 50 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab208182 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type cell lysates with no signal at this size in Smad3 knockout cell line ab263915 (knockout cell lysate ab264513). Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab208182 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-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 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD3 knockout A549 cell lysate (ab264513)

Lane 3:

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

Lane 4:

Human Kidney cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 50 kDa

false

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2 : SMAD3 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4 : Human Kidney cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab40854 observed at 48 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab40854 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type cell lysates with no signal at this size in Smad3 knockout cell line ab263915 (knockout cell lysate ab264513). Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab40854 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-Smad3 antibody [EP568Y] (<a href='/en-us/products/primary-antibodies/smad3-antibody-ep568y-ab40854'>ab40854</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD3 knockout A549 cell lysate (ab264513)

Lane 3:

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

Lane 4:

Human Kidney cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 50 kDa

false

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell lysate (AB264513)

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : SMAD3 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lanes 1 - 2 : Merged signal (red and green). Green - ab84177 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab84177 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type cell lysates with no signal at this size in Smad3 knockout cell line ab263915 (knockout cell lysate ab264513). Wild-type and SMAD3 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab84177 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml 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-Smad3 antibody (<a href='/en-us/products/primary-antibodies/smad3-antibody-ab84177'>ab84177</a>) at 1 µg/mL

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

SMAD3 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD3 knockout A549 cell lysate (ab264513)

Predicted band size: 48 kDa

Observed band size: 50 kDa

false

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 100%

Disease

Carcinoma

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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>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab264513-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human SMAD3 knockout A549 cell lysate", "number":"AB264513-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A549 cell lysate", "number":"AB264513-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
SMAD3
Gene editing type
Knockout
Gene editing method
CRISPR technology
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: 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
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