PA5-33559

antibody from Invitrogen Antibodies

Targeting: FZD7 FzE3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-33559

Provider

Invitrogen Antibodies

Product name

FZD7 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse, Rat, Bovine, Hamster, Porcine, Rabbit, Simian

Host

Rabbit

Isotype

IgG

Vial size

50 µg

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-FZD7 Polyclonal Antibody (Product # PA5-33559) and a 70 kDa band corresponding to Frizzled-7 was observed across the cell lines tested except SK-BR-3 which is reported to be negative. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), SW480 (Lane 2), HCT 116 (Lane 3), HT-29 (Lane 4), MDA-MB-231 (Lane 5) and SK-BR-3 (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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). 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

Immunofluorescence analysis of Frizzled-7 was performed using 70% confluent log phase MDA-MB-231 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 FZD7 Polyclonal Antibody (Product # PA5-33559) at 1:100 dilution 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 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: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma Membrane, Endoplasmic Reticulum and Cytoplasmic vesicular localization. Panel e represents SK-BR-3 cells having no expression of FZD7. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of formalin-fixed paraffin-embedded human heart, cardiac myocytes using a Frizzled 7/FZD7 polyclonal antibody (Product # PA5-33559). Heat-induced antigen retrieval was performed prior to staining.