PA5-18118

antibody from Invitrogen Antibodies

Targeting: NLRP3 AGTAVPRL, AII, AVP, C1orf7, CIAS1, CLR1.1, DFNA34, FCAS, FCU, MWS, NALP3, PYPAF1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-18118

Provider

Invitrogen Antibodies

Product name

NLRP3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine and rat based on sequence homology.

Reactivity

Human

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.5 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 NLRP3 Polyclonal Antibody (Product # PA5-18118) and a 120 kDa band corresponding to NLRP3 was observed only in THP-1 which is the positive cell model when compared to other cell lines tested along with uncharacterized bands (*) at ~40-50 kDa. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), SK-BR-3 (Lane 2), Raji (Lane 3), MDA-MB-231 (Lane 4), and SK-OV-3 (Lane 5) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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 NLRP3 in U937 cells using a NLRP3 monoclonal antibody (Product # PA5-18118) at 10 µg/mL for1hr. The cells were paraformaldehyde fixed and permeabilized with 0.15% Triton. Primary incubation was followed by Alexa Fluor 488 secondary antibody (4 µg/mL showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL)followed by Alexa Fluor 488 secondary antibody (4 µg/mL).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NLRP3 in U937 cells using a NLRP3 monoclonal antibody (Product # PA5-18118) at 10 µg/mL for1hr. The cells were paraformaldehyde fixed and permeabilized with 0.15% Triton. Primary incubation was followed by Alexa Fluor 488 secondary antibody (4 µg/mL showing cytoplasmic staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL)followed by Alexa Fluor 488 secondary antibody (4 µg/mL).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of NLRP3 in human peripheral blood monocytes using an NLRP3 monoclonal antibody (Product # PA5-18118) at 10 µg/mL for 1hr, depicted by the blue line. The cells were paraformaldehyde fixed and permeabilized with 0.5% Triton. Primary incubation followed by Alexa Fluor 488 secondary antibody (0.2 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of NLRP3 in U937 cells using a polyclonal antibody (Product #PA5-18118). U937 cells (blue line) were paraformaldehyde fixed and permeabilized with 0.5% Triton. The primary antibody was incubated for one hour (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (1 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by an Alexa Fluor 488 secondary antibody.