PA5-32366

antibody from Invitrogen Antibodies

Targeting: PTGS2 COX2

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-32366

Provider

Invitrogen Antibodies

Product name

COX2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is colon, lung or breast carcinoma.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Concentration

0.4 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-COX2 Rabbit Polyclonal Antibody (Product # PA5-32366) and a 74 kDa band corresponding to COX2 was induced in RAW 264.7 upon LPS treatment. Membrane enriched extracts (30 µg lysate) of RAW 264.7 (Lane 1) and RAW 264.7 treated with LPS (1µg/ml for 24 hr) (Lane 2) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris 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 (1:2000 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of COX2 was performed using 70% confluent log phase RAW 264.7 cells treated with 100 ng/mL of Lipopolysaccharide for 24 Hrs. 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 10 minutes at room temperature. The cells were labeled with COX2 Polyclonal Antibody (Product # PA5-32366) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034), (1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic and nuclear membrane localization. Panel e represents untreated cells with reduced signal. 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 COX-2 using anti-COX-2 Polyclonal Antibody (Product # PA5-32366) in Breast Carcinoma Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.