Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
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- Product number
- MA5-25574 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PTP1B Monoclonal Antibody (OTI1A2)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI1A2
- Vial size
- 100 µL
- Concentration
- 0.72 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PTPN1 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with PTPN1 (Product # MA5-25574) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PTPN1 in HeLa wild-type and knockout cells using 10 µg per lane. Samples were separated by SDS-PAGE and probed with PTPN1 (Product # MA5-25574) monoclonal antibody and HSP90AB1 antibody with a dilution of 1:500 as a loading control.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PTP1B was achieved by transfecting MCF-7 cells with PTP1B specific siRNAs (Silencer® select Product # s11508 ). Western blot analysis (Fig. a) was performed using membrane extracts from the PTP1B knockdown cells (Lane 2) and non-specific scrambled siRNA transfected cells (Lane 1). The blot was probed using PTP1B Monoclonal Antibody (Product # MA5-25574) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PTP1B.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PTP1B Monoclonal Antibody (Product # MA5-25574). A ~45kDa band corresponding to PTP1B was observed in Jurkat and MCF-7 cell lines. Membrane enriched extracts (30 µg) of Jurkat (Lane 1) and MCF-7 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of PTPN1 in COS7 cells. Cells were transfected with a plasmid overexpressing PTPN1 and probed with a PTPN1 monoclonal antibody (Product # MA5-25574).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PTP1B was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 60 minutes at room temperature. The cells were labeled with PTPB1 Monoclonal Antibody (Product # MA5-25574) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175), (1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.