Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸ 10 µl 110 563 ₸

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Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23094-1] to NLRP3
Suitable for: ELISA, WB, Flow Cyt, IP
Knockout validated
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-NLRP3 antibody [EPR23094-1]
See all NLRP3 primary antibodies
Description

Rabbit monoclonal [EPR23094-1] to NLRP3
Host species

Rabbit
Specificity

Stimulation may be required to allow detection of the NLRP3 protein in most samples.

Tested Applications & Species

Application	Species
ELISA	Recombinant fragment
Flow Cyt	Human
IP	Mouse Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: THP-1 and J774A.1 whole cell lysate. LPS treated RAW 264.7 whole cell lysate. Rat spleen lysate. Mouse Wt BMDCs treated with 50 ng/ml LPS, 3h; Human THP1 Wt cells treated with 200 ng/ml LPS, 3h. RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate. IP: RAW 264.7 and THP-1 whole cell lysate. Flow Cyt: THP-1 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23094-1
Isotype

IgG
Research areas

Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) This image is courtesy of Benedikt Saller, from Prof. Dr. Olaf Groß’ lab at the University of Freiburg.

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

Lane 1 : Mouse Wt BMDCs treated with 50 ng/ml LPS, 3h.
Lane 2 : Mouse NLRP3-/- BMDCs treated with 50 ng/ml LPS, 3h.
Lane 3 : Human THP1 Wt cells treated with 200 ng/ml LPS, 3h.

Lysates/proteins at 25000 cells per lane.

Secondary
All lanes : Goat anti-Rabbit HRP at 1/5000 dilution

Developed using the ECL technique.

Predicted band size: 118 kDa

Exposure time: 10 seconds

Blocking step: 2% SMP in PBS/T for 60 minutes at RT.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2 : Jurkat (human t cell leukemia t lymphocyte) whole cell lysate at 20 µg
Lane 3 : Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg
Lane 4 : RAW 264.7 treated with 10 µg/ml LPS for 8 hours, whole cell lysate at 10 µg
Lane 5 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 118 kDa

The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).

Note: The band around 75kDa is the short form of NLRP3 (NLRP3s).

Low expression cell line: Jurkat (PMID: 19767079).

Exposure times: Lane 1: 48 secs; Lane 2: 3 min; Lanes 3-4: 1 sec; Lane 5: 6 secs.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 118 kDa
Observed band size: 118 kDa

Exposure time: 15 seconds

Stimulation is required to allow detection of the NLRP3 protein in C6, PC-12 and NIH/3T3 cell lines.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution + Rat spleen lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 118 kDa

Exposure time: 3 minutes

The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

NLRP3 was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10µg with ab263899 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1: RAW 264.7 whole cell lysate 10µg

Lane 2: ab263899 IP in RAW 264.7 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263899 in RAW 264.7 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

NLRP3 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10µg with ab263899 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1: THP-1 whole cell lysate 10µg

Lane 2: ab263899 IP in THP-1 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263899 in THP-1 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 30 seconds.
Flow Cytometry - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labeling NLRP3 with ab263899 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.
ELISA - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

ELISA analysis of NLRP3 recombinant protein at 1000 ng/mL with ab263899. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Images

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