Invitrogen

NLRP3 Polyclonal Antibody

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NLRP3 Antibody in Immunocytochemistry (ICC/IF) — mmunofluorescent analysis of NLRP3 on THP-1 cells. Cell Climbing Slice Preparation: Cover slip with cells was washed 3X in PBS and then immersed in 4% paraformaldehyde or neutral formalin for 10-20 minutes. Cover slip was washed 3X in PBS and allowed to dry for 8-10 hours. The cells were blocked with 10% goat serum and then incubated with 2 µg/mL NLRP3 polyclonal antibody (Product# PA5-79740) overnight at 4°C. DyLight ®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody diluted 1:100 and incubated for 30 minutes at 37°C. Cells were visualized using a fluorescence microscope and filter sets appropriate for the label used.

NLRP3 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of NLRP3 on paraffin-embedded human tonsil tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NLRP3 polyclonal antibody (Product# PA5-79740) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

NLRP3 Antibody in Western Blot (WB) — Western blot analysis of NLRP3 in Lane 1: HEP-2 cell lysate, Lane 2: A549 cell lysate, Lane 3: U87 cell lysate, Lane 4: CEM cell lysate using 50 µg (reducing conditions) per well. Electrophoresis was performed on 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours and protein was transferred to a nitrocellulose membrane at 150mA for 50-90 minutes. Sample was blocked with 5% Non-fat Milk/TBS for 1.5 hours at room temperature, incubated with NLRP3 polyclonal antibody (Product # PA5-79740) at a dilution of 0.5 µg/mL (overnight at 4°C), followed by goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10,000. Signal development was performed using a chemiluminescence (ECL) kit.

NLRP3 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 3 SHED inhibited CCl 4 -induced NLRP3 inflammasome activation and pyroptosis in hepatocytes. (A-C) The mRNA level of Caspase-1, GSDMD, and IL-1b was detected after 3h and 6h CCl 4 stimulation analyzed by RT-qPCR. (D-H) The expression levels of NLRP3, Caspase-1, and GSDMD-C in hepatocytes in control, CCl 4 , and SHED infusion group by Western blotting analysis. (I-K) THe expression of NLRP3, Caspase-1 and GSDMD in hepatocytes in control, CCl 4 , and CCl 4 +SHED groups was analyzed by immunofluorescence staining. (L) The ratio of dead cells was determined by PI staining in control, CCl 4 , and CCl 4 +SHED groups. Scale bar: 50 mum. *p < 0.05, **p < 0.01, and ***p < 0.001. Graph bars show the mean +- SD. All of the assays were performed in triplicate. PI, propidium iodide; SHED, stem cells from human exfoliated deciduous teeth; GSDMD, gasdermin D.

NLRP3 Antibody in Immunohistochemistry (IHC) — Figure 2 SHED attenuated pyroptosis of hepatocytes in CCl 4 -induced liver cirrhosis. (A, B) Immunohistochemical staining of GSDMD in control, CCl 4 , and SHED transplantation groups (C-H) . The expression of NLRP3, Caspase-1, and IL-1beta in control, CCl 4 , and SHED transplantation groups, as assessed by immunohistochemical staining. Scale bar: 100 and 20 mum. *p < 0.05, **p < 0.01, and ***p < 0.001. Graph bars show the mean +- SD. All of the assays were performed in triplicate. NLRP3, NOD-like receptor family pyrin domain containing 3; GSDMD, gasdermin D; SHED, stem cells from human exfoliated deciduous teeth.

NLRP3 Antibody in Western Blot, Immunohistochemistry (WB, IHC) — The expression levels of NLRP3, ASC, caspase-1, and IL-18 in rat spinal cords 1, 3, 7, and 14 days after SCI. (A) Representative Western blotting and semiquantitative analysis of NLRP3 protein in the spinal cords of rats in the sham and model groups at days 1, 3, 7, and 14 post-SCI. The image shows representative Western blotting for NLRP3 and beta-actin. (B) Western blotting was used to analyze the protein expression of ASC in the sham and model groups after SCI. Representative protein bands are shown for ASC at 1, 3, 7, and 14 days after SCI (N = 5 rats/group). The expression of ASC is shown for the model and sham groups. *p < 0.05; two-way ANOVA. (C) The BBB function scores in the sham and model groups. N = 8 rats/group. *p < 0.05. (D) The expression of NLRP3 in the spinal cords of rats was detected using IF. The expression of NLRP3 in the spinal cord was detected in the sham group and 1, 3, and 7 days after SCI. Cells positive for NLRP3 protein are shown in green. (E) Quantitative analysis of NLRP3-positive cells in the model group compared with the sham group. *p < 0.05; one-way ANOVA was used for comparison. N = 5 rats/group. (F, G) Protein expression and representative protein bands for caspase-1 and IL-18 at each time point after SCI as determined using Western blotting. N = 5 rats/group. *p < 0.05; two-way ANOVA was used for comparison. Data are presented as the mean +- SEM. SCI, spinal cord injury; BBB, Basso-Beattie-Bresnahan; IF, immunofluorescence.

Product Details

PA5-79740

Applications	Tested Dilution	Publications
Western Blot (WB)	0.1-0.5 µg/mL	View 2 publications 2 publications
Immunohistochemistry (IHC)	-	View 3 publications 3 publications
Immunohistochemistry (Paraffin) (IHC (P))	0.5-1 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	2 µg/mL	View 1 publication 1 publication
Flow Cytometry (Flow)	1-3 µg/1x10^6 cells	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A synthetic peptide corresponding to a sequence at the N-terminus of human CIAS1 (12-31aa RYLEDLEDVDLKKFKMHLED).
Conjugate	Unconjugated
Form	Lyophilized
Concentration	500 µg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 4mg trehalose
Contains	no preservative
Storage conditions	-20°C
RRID	AB_2746855

Product Specific Information

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Target Information

This gene encodes a pyrin-like protein containing a pyrin domain, a nucleotide-binding site domain, and a leucine-rich repeat motif. This protein interacts with the apoptosis-associated speck-like protein PYCARD/ASC, which contains a caspase recruitment domain, and is a member of the NALP3 inflammasome complex. This complex functions as an upstream activator of NF-kappaB signaling, and it plays a role in the regulation of inflammation, the immune response, and apoptosis. Mutations in this gene are associated with familial cold autoinflammatory syndrome, and neonatal-onset multisystem inflammatory disease. Multiple alternatively spliced transcript variants encoding distinct isoforms have been identified for this gene. Alternative 5' UTR structures are suggested by available data; however, insufficient evidence is available to determine if all of the represented 5' UTR splice patterns are biologically valid.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Stem Cells From Human Exfoliated Deciduous Teeth Alleviate Liver Cirrhosis via Inhibition of Gasdermin D-Executed Hepatocyte Pyroptosis.

Frontiers in immunology

Chen P,Zhou YK,Han CS,Chen LJ,Wang YM,Zhuang ZM,Lin S,Zhou YH,Jiang JH,Yang RL

Published figure using NLRP3 polyclonal antibody (Product # PA5-79740) in Immunocytochemistry

Thu Jun 02 00:00:00 EDT 2022

Stem Cells From Human Exfoliated Deciduous Teeth Alleviate Liver Cirrhosis via Inhibition of Gasdermin D-Executed Hepatocyte Pyroptosis.

Frontiers in immunology

Chen P,Zhou YK,Han CS,Chen LJ,Wang YM,Zhuang ZM,Lin S,Zhou YH,Jiang JH,Yang RL

Published figure using NLRP3 polyclonal antibody (Product # PA5-79740) in Immunocytochemistry

Thu Jun 02 00:00:00 EDT 2022

Stem Cells From Human Exfoliated Deciduous Teeth Alleviate Liver Cirrhosis via Inhibition of Gasdermin D-Executed Hepatocyte Pyroptosis.

Frontiers in immunology

Chen P,Zhou YK,Han CS,Chen LJ,Wang YM,Zhuang ZM,Lin S,Zhou YH,Jiang JH,Yang RL

Published figure using NLRP3 polyclonal antibody (Product # PA5-79740) in Immunocytochemistry

Thu Jun 02 00:00:00 EDT 2022

Electroacupuncture Inhibits NLRP3 Activation by Regulating CMPK2 After Spinal Cord Injury.

Frontiers in immunology

Chen Y,Wu L,Shi M,Zeng D,Hu R,Wu X,Han S,He K,Xu H,Shao X,Ma R

Published figure using NLRP3 polyclonal antibody (Product # PA5-79740) in Immunohistochemistry

Tue Apr 12 00:00:00 EDT 2022

Electroacupuncture Inhibits NLRP3 Activation by Regulating CMPK2 After Spinal Cord Injury.

Frontiers in immunology

Chen Y,Wu L,Shi M,Zeng D,Hu R,Wu X,Han S,He K,Xu H,Shao X,Ma R

Published figure using NLRP3 polyclonal antibody (Product # PA5-79740) in Immunohistochemistry

Tue Apr 12 00:00:00 EDT 2022

NU9056, a KAT 5 Inhibitor, Treatment Alleviates Brain Dysfunction by Inhibiting NLRP3 Inflammasome Activation, Affecting Gut Microbiota, and Derived Metabolites in LPS-Treated Mice.

Frontiers in nutrition

Chen L,Qing W,Yi Z,Lin G,Peng Q,Zhou F

PA5-79740 was used in Immunohistochemistry (Paraffin) to investigate if NU9056 might effectively alleviate lipopolysaccharide-induced cognitive impairment and emotional disorder in experimental mice by inhibiting the NLRP3 inflammasome.

Sat Jul 31 00:00:00 EDT 2021

NLRP3 Inflammasome: A Potential Target in Isoflurane Pretreatment Alleviates Stroke-Induced Retinal Injury in Diabetes.

Frontiers in cellular neuroscience

Lin HB,Lin YH,Zhang JY,Guo WJ,Ovcjak A,You ZJ,Feng ZP,Sun HS,Li FX,Zhang HF

PA5-79740 was used in Immunohistochemistry to investigate whether isoflurane offers retinal protection against stroke-induced retinal injury in diabetes.

Tue Jul 27 00:00:00 EDT 2021

NLRP3 Inflammasome: A Potential Target in Isoflurane Pretreatment Alleviates Stroke-Induced Retinal Injury in Diabetes.

Frontiers in cellular neuroscience

Lin HB,Lin YH,Zhang JY,Guo WJ,Ovcjak A,You ZJ,Feng ZP,Sun HS,Li FX,Zhang HF

PA5-79740 was used in Immunohistochemistry to investigate whether isoflurane offers retinal protection against stroke-induced retinal injury in diabetes.

Tue Jul 27 00:00:00 EDT 2021

Bioinformatics

Protein Aliases: Angiotensin/vasopressin receptor AII/AVP-like; Caterpiller protein 1.1; CLR1.1; cold autoinflammatory syndrome 1 homolog; cold autoinflammatory syndrome 1 protein; Cold autoinflammatory syndrome 1 protein homolog; Cold-induced autoinflammatory syndrome 1 protein; Cryopyrin; FLJ95925; mast cell maturation inducible protein 1; Mast cell maturation-associated-inducible protein 1; NACHT domain-, leucine-rich repeat-, and PYD-containing protein 3; NACHT, LRR and PYD containing protein 3; NACHT, LRR and PYD domains-containing protein 3; NACHT/LRR/pyrin domain-containing protein 3; NLR family, pyrin domain containing 3; nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3; PYRIN-containing APAF1-like protein 1

Gene Aliases: AGTAVPRL; AII; AII/AVP; AVP; C1orf7; CIAS1; CLR1.1; FCAS; FCAS1; FCU; Mmig1; MWS; NALP3; NLRP3; PYPAF1

UniProt ID: (Human) Q96P20, (Rat) D4A523, (Mouse) Q8R4B8

Entrez Gene ID: (Human) 114548, (Rat) 287362, (Mouse) 216799

Molecular Function: non-motor microtubule binding protein