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AB255430

Human SMAD2 knockout HeLa cell line

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SMAD2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 이미지
Western blot - Human SMAD2 knockout HeLa cell line (AB255430)
  • WB

Lab

Western blot - Human SMAD2 knockout HeLa cell line (AB255430)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

Lanes 1-4 : Merged signal (red and green). Green - ab280888 observed at 60 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

ab280888 Anti-Smad2 (phospho S467) antibody [EPR23681-40] was shown to specifically react with Smad2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255430 (knockout cell lysate ab263833) was used. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE.

ab280888 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Smad2 (phospho S467) antibody [EPR23681-40] (<a href='/ko/products/primary-antibodies/smad2-phospho-s467-antibody-epr23681-40-ab280888'>ab280888</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) treated with 20 ng/ml TGF beta1 for 15 minutes, whole cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD2 knockout HeLa cell line (ab255430)

Lane 3:

Smad2 knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

Smad2 knockout HeLa (human cervix adenocarcinoma epithelial cell), treated with 20 ng/ml TGF beta1 for 15 minutes, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 60 kDa

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Western blot - Human SMAD2 knockout HeLa cell line (AB255430)
  • WB

Supplier Data

Western blot - Human SMAD2 knockout HeLa cell line (AB255430)

Lanes 1 - 4 : Merged signal (red and green). Green - ab40855 observed at 58 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab40855 was shown to react with Smad2 in wild-type HeLa. Loss of signal was observed when knockout cell line ab255430 (knockout cell lysate ab263833) was used. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. ab40855 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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-Smad2 antibody [EP784Y] (<a href='/ko/products/primary-antibodies/smad2-antibody-ep784y-ab40855'>ab40855</a>) at 1/2000 dilution

Lane 1:

A549 cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD2 knockout HeLa cell line (ab255430)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

SMAD2 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 52 kDa

Observed band size: 37 kDa,55 kDa

false

Western blot - Human SMAD2 knockout HeLa cell line (AB255430)
  • WB

Lab

Western blot - Human SMAD2 knockout HeLa cell line (AB255430)

Lanes 1 - 4 : Merged signal (red and green). Green - ab33875 observed at 58 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab33875 was shown to react with Smad2 in wild-type HeLa. Loss of signal was observed when knockout cell line ab255430 (knockout cell lysate ab263833) was used. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. ab33875 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-Smad2 antibody [EP567Y] (<a href='/ko/products/primary-antibodies/smad2-antibody-ep567y-ab33875'>ab33875</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human SMAD2 knockout HeLa cell line (ab255430)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

SMAD2 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 52 kDa

Observed band size: 37 kDa,58 kDa

false

Sanger Sequencing - Human SMAD2 knockout HeLa cell line (AB255430)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMAD2 knockout HeLa cell line (AB255430)

Allele-2 : 1 bp insertion in exon 2.

Sanger Sequencing - Human SMAD2 knockout HeLa cell line (AB255430)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMAD2 knockout HeLa cell line (AB255430)

Allele-1 : 1 bp deletion in exon 2.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2

질병

Adenocarcinoma

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보완 제품

Cell Lines & Lysates

AB255431

Human SMAD3 knockout HeLa cell line

Advanced Validation

Western blot - Human SMAD3 knockout HeLa cell line (AB255431)

  • 0
  • 0
Cell Lines & Lysates

AB255439

Human TGFB1 (TGF beta 1) knockout HeLa cell line

Advanced Validation

Western blot - Human TGFB1 (TGF beta 1) knockout HeLa cell line (AB255439)

  • 0
  • 0
Cell Lines & Lysates

AB276092

Human TP53 (p53) knockout A549 cell line

Advanced Validation

Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)

  • 0
  • 0
Primary Antibodies

AB6276

Anti-beta Actin antibody [AC-15] - Loading Control

KO Validated|Lab Essentials

Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (AB6276)

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Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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제품 세부 정보

We will provide viable cells that proliferate on revival.

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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제품 구성

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특성 및 보관 정보

유전자명
SMAD2
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
품질 관리 - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C
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추가 정보

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

Smad2 also known as Mothers against decapentaplegic homolog 2 (MAD2) or MADR2 is a signaling protein involved in the transforming growth factor-beta (TGF-β) receptor pathway. Smad2 has a molecular mass of approximately 58 kDa and expresses in various tissues including epithelial mesenchymal and endothelial cells. Smad2 undergoes phosphorylation on serine residues in response to TGF-β signaling converting it into phosphorylated forms often referred to as p-Smad2 or phospho-Smad2. These phosphorylated forms are critical for the relay of signals from the cell surface to the nucleus.
Biological function summary

Smad2 acts as an intracellular mediator for TGF-β signaling a pathway important for regulating cell proliferation differentiation and apoptosis. Smad2 typically functions as part of a heteromeric complex with Smad4 another key player in TGF-β signaling. Upon activation phosphorylated Smad2 combines with Smad4 to form a complex that translocates into the nucleus. This complex then binds to specific DNA sequences to regulate the transcription of target genes involved in processes such as cell growth inhibition and extracellular matrix production.

Pathways

Smad2 is integral to the TGF-β and activin receptor signaling pathways. These pathways are essential in controlling cell growth and immune responses. Smad2 interacts with other proteins such as Smad3 in addition to Smad4 to modulate gene expression effectively. The interaction between Smad2 and these proteins ensures precise control of cellular responses to external stimuli emphasizing its pivotal role in maintaining cellular homeostasis.

Smad2 correlates with various pathological conditions including fibrosis and cancer. Aberrant Smad2 signaling can contribute to the development of these diseases as excessive TGF-β signaling promotes fibrotic tissue deposition and tumor progression. Smad2 connects with other proteins like Smad3 in these pathological contexts both acting as mediators of abnormal cellular behaviors. Understanding the regulatory mechanisms of Smad2 can help develop therapeutic strategies against disorders linked to dysregulated TGF-β signaling.
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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

초기 취급 지침

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

Subculture 가이드라인
  • 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.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

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

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제품 프로토콜

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타겟 정보

See full target information SMAD2

대체 명칭 보기

SMAD 2, Mothers against DPP homolog 2, SMAD2, MAD homolog 2, Mad-related protein 2, hMAD-2, JV18-1, Smad2, MADR2, SMAD family member 2, Mothers against decapentaplegic homolog 2, MADH2, hSMAD2
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Product promise

저희는 고품질 시약으로 고객님의 연구를 서포트하고자 최선을 다하고 있으며, 모든 단계에서 함께 하겠습니다. 만약 제품이 기대한 성능을 보이지 않을 경우에도 Product Promise로 보호받으실 수 있습니다.
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