PA5-66880

antibody from Invitrogen Antibodies

Targeting: FOSL1 fra-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-66880

Provider

Invitrogen Antibodies

Product name

Fra1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: LVPSINTMSGS QELQWMVQPH FLGPSSYPRP LTYPQYSPPQ PRPGVIRALG PPPGVRRRPC EQISPE

Concentration

0.2 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Fra1 Polyclonal Antibody (Product # PA5-66880) and 30-40k Da bands corresponding to Fra1 were observed across cell lines tested. Nuclear enriched extracts (30 µg lysate) of PC-3 (Lane 1), LNCaP (Lane 2), U-2 OS (Lane 3), SH-SY5Y (Lane 4), IMR-32 (Lane 5) or HEK-293 (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:250) 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

Immunofluorescent staining of Fra1 in human cell line U-251 MG shows localization to nucleoplasm. Samples were probed using a Fra1 Polyclonal Antibody (Product # PA5-66880).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Fra1 was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Fra1 Polyclonal Antibody (Product # PA5-66880) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2500), 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 localization. Panel e shows merged image for HEK-293 cells which show no expression of the target protein. Panel f represents control PC-3 cells with no primary antibody to assess background. The images were captured at 63X magnification.