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AB257618

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

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PTPN11 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and 2 bp deletion in exon1.

View Alternative Names

BPTP3, JMML, METCDS, MGC14433, OTTHUMP00000166107, OTTHUMP00000166108, PTN11_HUMAN, PTP-1D, PTP-2C, PTPN11, Protein tyrosine phosphatase 2, Protein tyrosine phosphatase non receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SAP-2, SH-PTP2, SH-PTP3, SH2 domain containing protein tyrosine phosphatase 2, SHP-2, Tyrosine-protein phosphatase non-receptor type 11

4 Images
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)
  • WB

Lab

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : PTPN11 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab187040 observed at 68 kDa. Red - loading control ab8245 observed at 37 kDa.

ab187040 Anti-SHP2 antibody [EPR17829-9] was shown to specifically 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 and SHP2 knockout samples 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) were incubated overnight at 4°C at 1 in 5000 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-SHP2 antibody [EPR17829-9] (<a href='/en-us/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 (<a href='/en-us/products/cell-lines/human-ptpn11-shp2-knockout-hek-293t-cell-line-ab266450'>ab266450</a>)

Predicted band size: 68 kDa

Observed band size: 68 kDa

false

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)
  • WB

Lab

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : PTPN11 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab32083 observed at 68 kDa. Red - loading control ab8245 observed at 37 kDa.

ab32083 Anti-SHP2 antibody [Y478] was shown to specifically 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 and SHP2 knockout samples 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) 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-SHP2 antibody [Y478] (<a href='/en-us/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 (<a href='/en-us/products/cell-lines/human-ptpn11-shp2-knockout-hek-293t-cell-line-ab266450'>ab266450</a>)

Predicted band size: 68 kDa

Observed band size: 68 kDa

false

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)

Allele-1 : 2 bp deletion in exon1

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell lysate (AB257618)

Allele-2 : 1 bp insertion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and 2 bp deletion in exon1.

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Reactivity data

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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
PTPN11
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

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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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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Complementary Products

Primary Antibodies

AB32083

Anti-SHP2 antibody [Y478]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP2 antibody [Y478] (AB32083)

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Primary Antibodies

AB199376

Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free)

RabMAb|Advanced Validation|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (AB199376)

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Primary Antibodies

AB176094

Rabbit IgG, polyclonal - Isotype Control (Low Endotoxin, Azide-Free)

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Primary Antibodies

AB172730

Rabbit IgG, monoclonal [EPR25A] - Isotype Control

RabMAb|Advanced Validation|Recombinant

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

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Product promise

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