PA5-27803

antibody from Invitrogen Antibodies

Targeting: PTPN6 HCP, HCPH, PTP-1C, SHP-1, SHP1

Antibody data

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Validation data

Product number: PA5-27803 - Provider product page
Provider: Invitrogen Antibodies
Product name: SHP-1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1.29 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

PTPN6 protein structure - PA5-27803 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SHP1 using 30 µg of A431 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a SHP1 polyclonal antibody (Product # PA5-27803) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SHP-1 Polyclonal Antibody (Product # PA5-27803). Sample (30 µg of whole cell lysate). A: NIH-3T3. 7.5% SDS PAGE. SHP-1 Polyclonal Antibody (Product # PA5-27803) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SHP-1 Polyclonal Antibody (Product # PA5-27803). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with SHP-1 Polyclonal Antibody (Product # PA5-27803) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SHP-1 Polyclonal Antibody (Product # PA5-27803). Various tissue extracts (50 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with SHP-1 Polyclonal Antibody (Product # PA5-27803) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-SHP-1 Polyclonal Antibody(Product # PA5-27803) and a 63kDa band corresponding to SHP-1 was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of Jurkat (Lane 1), THP-1 (Lane 2), HEL 92.1.7 (Lane 3), K-562 (Lane 4), U-2 OS (Lane 5), Mouse Spleen (Lane 6) and Mouse Skeletal Muscle (Lane 7), were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).Jurkat, THP-1, HEL 92.1.7 and Spleen are expected to be high expressing cell lines and tissue, whereas, U-2 OS and skeletal muscle are expected to be low expressing cell line and tissue, as reported.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin immunoprecipitation analysis of PPM1A was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a PPM1A polyclonal antibody (Product # PA5-29041). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify exon 3-3 of human Bcl-x (hBcl-x) or exon 1 of human NR4A3 (hNR4A3). PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. Schematic representations of the Bcl-x and NR4A3 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions). Regions amplified by Bcl-x and NR4A3 primers are represented by black bars. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin immunoprecipitation analysis of SHP1 was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a SHP1 polyclonal antibody (Product # PA5-27803). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify exon 3-3 of human Bcl-x (hBcl-x) or exon 1 of human NR4A3 (hNR4A3). PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. Schematic representations of the Bcl-x and NR4A3 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions). Regions amplified by Bcl-x and NR4A3 primers are represented by black bars. Data courtesy of the Innovators Program.