PA1-12376

antibody from Invitrogen Antibodies

Targeting: PPP1CA PP-1A, PP1A, PP1alpha, PPP1A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

PA1-12376

Provider

Invitrogen Antibodies

Product name

PP1 alpha Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody recognizes the recombinant pp1 alpha, pp1 beta, pp1 gamma 1, and pp1 gamma 2 forms.

Reactivity

Human, Mouse, Rat, Bovine

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-PP1 alpha Polyclonal Antibody(Product # PA1-12376) and a 38kDa band corresponding to PP1 alpha was observed across all tested cell lines and tissues. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), A549 (Lane 3), PC-3 (Lane 4), Hep G2 (Lane 5), T98G (Lane 6), HEL 92.1.7 (Lane 7), Raji (Lane 8), THP-1 (Lane 9), Mouse Spleen (Lane 10) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of PP1 alpha was performed using 70 percent confluent log phase HeLa 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 45 minutes at room temperature. The cells were labeled with PP1 alpha Polyclonal Antibody (Product # PA1-12376) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.