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AB266450

Human PTPN11 (SHP2) knockout HEK-293T cell line

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PTPN11 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 insertion in exon 1 and 2 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

대체 명칭 보기

SH-PTP3, PTPN11, SHPTP2, SH-PTP2, Tyrosine-protein phosphatase non-receptor type 11, PTP2C, Protein-tyrosine phosphatase 2C, PTP-1D, Protein-tyrosine phosphatase 1D, Shp2, PTP-2C, SHP-2

6 이미지
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • WB

Lab

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Lanes 1- 2 : Merged signal (red and green). Green - ab187040 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab187040 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266450 (knockout cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187040 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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-SHP2 antibody [EPR17829-9] (<a href='/ko/products/primary-antibodies/shp2-antibody-epr17829-9-ab187040'>ab187040</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PTPN11 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (ab266450)

Predicted band size: 68 kDa

Observed band size: 68 kDa

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Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • WB

Lab

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Lanes 1- 2 : Merged signal (red and green). Green - ab32083 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32083 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266450 (knockout cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-SHP2 antibody [Y478] (<a href='/ko/products/primary-antibodies/shp2-antibody-y478-ab32083'>ab32083</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PTPN11 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (ab266450)

Predicted band size: 68 kDa

Observed band size: 68 kDa

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Cell Culture - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • Cell Culture

Lab

Cell Culture - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Representative images PTPN11 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Allele-1 : 2 bp deletion in exon1

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Allele-2 : 1 bp insertion in exon 1.

Immunocytochemistry/ Immunofluorescence - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PTPN11 KO HEK293T (PTPN11 KO knock out human embryonic kidney epithelial cell) (ab266450) cells labelling SHP2 with ab300579 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in wildtype HEK293T cell line, while showing no staining in PTPN11 knockout HEK293T cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

주요 정보

세포 유형

HEK-293T

Species or organism

Human

조직

Kidney

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

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

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보완 제품

Biochemicals

AB120751

Thioflavin T, Fluorescent cell-permeable amyloid binding benzothiazole salt

Chemical Structure - Thioflavin T, Fluorescent cell-permeable amyloid binding benzothiazole salt (AB120751)

  • 0
  • 0
Assay Kits

AB260052

Human Alpha-synuclein ELISA Kit

KO Validated|Recombinant|SimpleStep

ELISA - Human Alpha-synuclein ELISA Kit (AB260052)

  • 5
  • 2
Primary Antibodies

AB136333

PE Anti-DC-SIGN antibody [UW60.1]

Flow Cytometry - PE Anti-DC-SIGN antibody [UW60.1] (AB136333)

  • 0
  • 0
Primary Antibodies

AB51249

Anti-Presenilin 2/AD5 antibody [EP1515Y]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

  • 4
  • 7
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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
PTPN11
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
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.

SHP2 also known as PTPN11 is a protein tyrosine phosphatase with a molecular mass of approximately 68 kDa. It is expressed in various tissues including the heart liver and immune cells. SHP2 belongs to the non-receptor class of protein tyrosine phosphatases and plays a critical role in cell signaling by acting as a regulator of signal transduction processes. SHP2 mediates these processes by dephosphorylating specific phosphotyrosine residues on target proteins influencing various cellular functions like proliferation differentiation and survival.
Biological function summary

The role of SHP2 extends to involvement in several signaling cascades such as the Ras/MAPK and PI3K/AKT pathways. It functions as an essential component within protein complexes that facilitate cell communication and response to external signals. The protein modulates growth factor signaling and cytokine signaling highlighting its significance in normal cell function and development. SHP2's statement in signaling processes makes it an important regulator of cellular dynamics.

Pathways

SHP2 participates in the Ras/MAPK and PI3K/AKT signaling pathways which are important for regulating cell growth survival and differentiation. Within these pathways SHP2 interacts with various signaling molecules including Grb2 Sos and Gab family adaptors. These interactions coordinate cellular responses to growth factors and other extracellular cues ensuring proper pathway activation and control. By serving as a critical mediator SHP2 integrates signals that are necessary for appropriate cellular outcomes.

SHP2 is associated with several conditions such as Noonan syndrome and various cancers. Mutations in the PTPN11 gene which encodes SHP2 often result in aberrant signaling that leads to developmental anomalies or tumorigenesis. In Noonan syndrome the mutated SHP2 protein results in disrupted Ras/MAPK pathway signaling. As for cancers SHP2 is often found to be overactive leading to enhanced cell proliferation and survival. In these contexts SHP2 is interconnected with other proteins like RAS and RAF which also contribute to oncogenic pathway activation and disease progression.
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품질 관리

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

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 PTPN11
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Product promise

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