PA5-52006

antibody from Invitrogen Antibodies

Targeting: FOXP1 12CC4, hFKH1B, HSPC215, QRF1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-52006

Provider

Invitrogen Antibodies

Product name

FOXP1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: QMQQLQQQHL LSLQRQGLLT IQPGQPALPL QPLAQGMIPT ELQQLWKEVT SAHTAEETTG NNHSSLDLTT TCVSSSAPSK TSLIMNPHAS TNGQLSVHTP KRESLSHEEH PHSHPLYGHG VCKWPGCEAV CEDFQSFLKH LNS

Concentration

0.15 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FOXP1 in Lane 1: NIH-3T3 cell lysate (Mouse embryonic fibroblast cells); Lane 2: NBT-II cell lysate (Rat Wistar bladder tumour cells). Samples were probed using a FOXP1 Polyclonal Antibody (Product # PA5-52006).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-FOXP1 Polyclonal Antibody (Product # PA5-52006) and 50-80 kDa bands corresponding to FOXP1 were observed across cell lines and tissues tested. Whole cell or tissue extracts (30 µg lysate) of Reh (Lane 1), Daudi (Lane 2), Jurkat (Lane 3), Hep G2 (Lane 4), BeWo (Lane 5), THP-1 (Lane 6), Mouse Lung (Lane 7), Mouse Spleen (Lane 8) or Mouse Liver (Lane 9) 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 (1:500) 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 FOXP1 in human cell line U-251 MG shows positivity in nucleus but excluded from the nucleoli. Samples were probed using a FOXP1 Polyclonal Antibody (Product # PA5-52006).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of FOXP1 was performed using 70% confluent log phase Hep G2 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 FOXP1 Polyclonal Antibody (Product # PA5-52006) at 1:200 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), (1:2500), 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 represents control cells with no primary antibody to assess background. The images were captured at 63X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of FOXP1 in human tonsil using a FOXP1 Polyclonal Antibody (Product # PA5-52006) shows moderate nuclear positivity in non-germinal center cells.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous FOXP1 protein using FOXP1 Antibody: ChIP was performed using Anti-FOXP1 Polyclonal Antibody (Product # PA5-52006, 2.5 µg) on sheared chromatin from LnCaP cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to PSA, and HSP90AB1 (Active) and SAT2 satellite repeats (Inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.