MA1-23236

antibody from Invitrogen Antibodies

Targeting: FOXO1 FKH1, FKHR, FOXO1A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-23236

Provider

Invitrogen Antibodies

Product name

FOXO1 Monoclonal Antibody (7H3)

Antibody type

Monoclonal

Antigen

Other

Description

Predicted molecular weight of approximately 70 kDa. Positive control of Cos 1 cells transfected with human FKHR suggested.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

7H3

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-FOXO1 Monoclonal Antibody (7H3) (Product # MA1-23236) and a ~80 kDa band corresponding to Forkhead box protein O1 was observed in REH and DAUDI, and was absent in A375 and Jurkat; along with an uncharacterised band (*) at ~40 kDa across cell lines tested. Whole cell extracts (30 µg lysate) of REH (Lane 1), DAUDI (Lane 2), A375 (Lane 3) and Jurkat (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:20000 dilution using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of FOXO1 in paraformaldehyde-fixed HeLa cells using a FOXO1 monoclonal antibody (Product # MA1-23236) at a 1:100 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Forkhead box protein O1 was performed using 70% confluent log phase Reh 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 overnight at room temperature. The cells were labeled with FOXO1 Monoclonal Antibody (14E4) (Product # MA1-23236) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), 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 Cytoplasmic localization. Panel e represents Jurkat cells having no expression of FOXO1. Panel f represent control cells with no primary antibody to assess background. The images were captured at 60x magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using FOXO1 (Product # MA1-23236) antibody at 1:100 dilution.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded Colon ca, using FOXO1 (Product # MA1-23236) antibody at 1:50 dilution.