PA1-37469

antibody from Invitrogen Antibodies

Targeting: CLDN3 C7orf1, CPE-R2, CPETR2, HRVP1, RVP1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-37469

Provider

Invitrogen Antibodies

Product name

Claudin 3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, canine, mouse, porcine and rat based on sequence homology. 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 small intestine tissue.

Reactivity

Human, Mouse, Rat, Chicken/Avian, Porcine

Host

Rabbit

Isotype

IgG

Vial size

1 mL

Storage

-20°C or -80°C if preferred

References

Submitted references

Disruption of the pro-inflammatory, anti-inflammatory cytokines and tight junction proteins expression, associated with changes of the composition of the gut microbiota in patients with irritable bowel syndrome.

Ivashkin V, Poluektov Y, Kogan E, Shifrin O, Sheptulin A, Kovaleva A, Kurbatova A, Krasnov G, Poluektova E
PloS one 2021;16(6):e0252930

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Claudin 3 Polyclonal Antibody (Product # PA1-37469, PA5-32353) and a 20 kDa band corresponding to Claudin-3 was observed across tested samples. Membrane enriched extracts (40 µg lysate) of MCF7 (Lane 1), HT-29 (Lane 2), SK-O-V3 (Lane 3), NIH:OVCAR-3 (Lane 4), U-937 (Lane 5), SK-BR-3 (Lane 6), U-87 MG (Lane 7) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Claudin-3 was performed using 70% confluent log phase NIH:OVCAR-3 cells. The cells were fixed with 4% paraformaldehyde for 5 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 Claudin 3 Polyclonal Antibody (Product # PA1-37469, PA5-32353) 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: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell junctions and nucleus localization. Panel e represents low expressing SK-OV-3 cells. 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 Claudin-3 using anti-Claudin-3 Polyclonal Antibody (Product # PA5-32353) in Small Intestine Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of Claudin 3 using a polyclonal antibody (Product # PA1-37469).