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Anti-SHP2/PTPN11 Antibody Picoband™ (monoclonal, 2E6)

Figure 1. Western blot analysis of SHP2/PTPN11 using anti-SHP2/PTPN11 antibody (M00150-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human CCRF-CEM whole cell lysates.
Lane 4: human K562 whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human CACO-2 whole cell lysates,
Lane 7: human Hela whole cell lysates,
Lane 8: rat brain tissue lysates,
Lane 9: rat C6 whole cell lysates,
Lane 10: mouse brain tissue lysates,
Lane 11: mouse Neuro-2a whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-SHP2/PTPN11 antigen affinity purified monoclonal antibody (Catalog # M00150-2) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for SHP2/PTPN11 at approximately 70KD. The expected band size for SHP2/PTPN11 is at 70KD.
Figure 2. IHC analysis of SHP2/PTPN11 using anti SHP2/PTPN11 antibody (M00150-2).
SHP2/PTPN11 was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-SHP2/PTPN11 Antibody (M00150-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 3. IHC analysis of SHP2/PTPN11 using anti SHP2/PTPN11 antibody (M00150-2).
SHP2/PTPN11 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-SHP2/PTPN11 Antibody (M00150-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 4. IHC analysis of SHP2/PTPN11 using anti SHP2/PTPN11 antibody (M00150-2).
SHP2/PTPN11 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-SHP2/PTPN11 Antibody (M00150-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 5. IHC analysis of SHP2/PTPN11 using anti SHP2/PTPN11 antibody (M00150-2).
SHP2/PTPN11 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-SHP2/PTPN11 Antibody (M00150-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 6. IF analysis of SHP2/PTPN11 using anti-SHP2/PTPN11 antibody (M00150-2).
SHP2/PTPN11 was detected in immunocytochemical section of U251 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-SHP2/PTPN11 Antibody (M00150-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 7. Flow Cytometry analysis of A549 cells using anti-SHP2/PTPN11 antibody (M00150-2).
Overlay histogram showing A549 cells stained with M00150-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-SHP2/PTPN11 Antibody (M00150-2, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	Q06124
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-SHP2/PTPN11 Antibody Picoband™ (monoclonal, 2E6) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Add 0.2ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	5781
Other Names	Tyrosine-protein phosphatase non-receptor type 11, 3.1.3.48, Protein-tyrosine phosphatase 1D, PTP-1D, Protein-tyrosine phosphatase 2C, PTP-2C, SH-PTP2, SHP-2, Shp2, SH-PTP3, PTPN11, PTP2C, SHPTP2
Calculated MW	68011 Da
Application Details	Western blot, 0.1-0.5 µg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml Immunocytochemistry/Immunofluorescence, 2 µg/ml Flow Cytometry, 1-3 µg/1x10^6 cells
Subcellular Localization	Nucleus. Cytoplasm.
Tissue Specificity	Widely expressed, with highest levels in heart, brain, and skeletal muscle.
Source	Eukaryota
Contents	Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clone Names	Clone: Clone: 2E6
Immunogen	A synthetic peptide corresponding to a sequence at the N-terminus of human SHP2, identical to the related mouse and rat sequences.
Cross Reactivity	No cross-reactivity with other proteins.
Storage	Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	PTPN11
Synonyms	PTP2C, SHPTP2
Function	Acts downstream of various receptor and cytoplasmic protein tyrosine kinases to participate in the signal transduction from the cell surface to the nucleus (PubMed:10655584, PubMed:14739280, PubMed:18559669, PubMed:18829466, PubMed:26742426, PubMed:28074573, PubMed:32184441). Positively regulates MAPK signal transduction pathway (PubMed:28074573). Dephosphorylates GAB1, ARHGAP35 and EGFR (PubMed:28074573). Dephosphorylates ROCK2 at 'Tyr-722' resulting in stimulation of its RhoA binding activity (PubMed:18559669). Dephosphorylates CDC73 (PubMed:26742426). Dephosphorylates SOX9 on tyrosine residues, leading to inactivate SOX9 and promote ossification (By similarity). Dephosphorylates tyrosine-phosphorylated NEDD9/CAS-L (PubMed:19275884). Acts as an effector of PDCD1-mediated inhibition of T-cell response: recruited by phosphorylated PDCD1, mediating dephosphorylation of key T-cell receptor (TCR) proximal signaling molecules, leading to TCR signaling inhibition (PubMed:32184441).
Cellular Location	Cytoplasm. Nucleus
Tissue Location	Widely expressed, with highest levels in heart, brain, and skeletal muscle.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

PTPN11 (Tyrosine-protein phosphatase non-receptor type 11), also known as protein-tyrosine phosphatase 1D (PTP-1D), protein-tyrosine phosphatase 2C (PTP-2C), TYROSINE PHOSPHATASE SHP2 (SHP2), BPTP3, SH-PTP2, SHP-2, SH-PTP3, is an enzyme that in humans is encoded by the PTPN11 gene. PTPN11 is a member of the protein tyrosine phosphatase (PTP) family. The open reading frame consists of 1,779 nucleotides potentially encoding a protein of 593 amino acids with a predicted molecular mass of 68 kD. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. This PTP contains two tandem Src homology-2 domains, which function as phospho-tyrosine binding domains and mediate the interaction of this PTP with its substrates. This PTP is widely expressed in most tissues and plays a regulatory role in various cell signaling events that are important for a diversity of cell functions, such as mitogenic activation, metabolic control, transcription regulation, and cell migration. Mutations in this gene are a cause of Noonan syndrome as well as acute myeloid leukemia.

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¥ 3,367.00

Cat# ABO14824

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Anti-SHP2/PTPN11 Antibody Picoband™ (monoclonal, 2E6)

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