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AB263915

Human SMAD3 knockout A549 cell line

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SMAD3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human SMAD3 knockout A549 cell line (AB263915)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell line (AB263915)

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 react with Smad3 in wild-type A549 cells in western blot with loss of signal observed in SMAD3 knockout sample. 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 (<a href='/en-us/products/cell-lysates/human-smad3-knockout-a549-cell-lysate-ab264513'>ab264513</a>)

Predicted band size: 48 kDa

Observed band size: 50 kDa

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Western blot - Human SMAD3 knockout A549 cell line (AB263915)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell line (AB263915)

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 react with Smad3 in western blot. 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 (<a href='/en-us/products/cell-lysates/human-smad3-knockout-a549-cell-lysate-ab264513'>ab264513</a>)

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 line (AB263915)
  • WB

Lab

Western blot - Human SMAD3 knockout A549 cell line (AB263915)

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 react with Smad3 in western blot. 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 (<a href='/en-us/products/cell-lysates/human-smad3-knockout-a549-cell-lysate-ab264513'>ab264513</a>)

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

Next Generation Sequencing - Human SMAD3 knockout A549 cell line (AB263915)
  • NGS

Supplier Data

Next Generation Sequencing - Human SMAD3 knockout A549 cell line (AB263915)

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

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

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

We will provide viable cells that proliferate on revival.

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

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?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab263915-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab259777 Human wild-type A549 cell line", "number":"AB263915-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab263915 Human SMAD3 knockout A549 cell line", "number":"AB263915-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab263915-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab263915 Human SMAD3 knockout A549 cell line", "number":"AB263915-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
SMAD3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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Target data

See full target information SMAD3
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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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