LF-MA0212

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

LF-MA0212

Provider

Invitrogen Antibodies

Product name

Anti-SHP-1 Monoclonal Antibody (14D5)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

A suggested positive control for this product is HL-60 cells.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

14D5

Vial size

100 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of Raji (Lane 1), Jurkat (Lane 2), MCF7 (Lane 3) and Caco-2 (Lane 4). The blot was probed with Anti-SHP-1 Mouse Monoclonal Antibody (Product # LF-MA0212, 1:2,000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 67 kDa band corresponding to SHP-1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SHP-1 Monoclonal Antibody was performed using 70% confluent log phase K562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SHP-1 (14D5) Mouse Monoclonal Antibody (Product # LF-MA0212) at 1:250 dilution in 0.1% BSA, incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 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 and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification