NALP4 Antibody - BSA Free

Immunohistochemistry: NALP4 Antibody [NB100-56156] - Formalin-fixed, paraffin-embedded tissue cores from a human breast cancer microarray stained for Smac/Diablo using NB100-56156 at 1:2000. Hematoxylin-eosin counterstain. Differential Smac/DIABLO staining is seen between patient samples.

NLRP4 expression induced olaparib resistance in pancreatic cancer cells.a, b MTS assays were performed to calculate the IC50 of olaparib in the indicated cells. c, d The indicated cells were treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). MTS assays were performed to investigate the effect of NLRP4 on olaparib resistance. The analysis of significant differences was performed with one-way ANOVA. ***p < 0.001. e The indicated cells were treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). Colony formation assays were performed to investigate the effect of NLRP4 on olaparib resistance. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. **p < 0.01. f, g Flow cytometry results for annexin V/7-ADD staining in the indicated cell lines after exposure to DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). The analysis of significant differences was performed with Student’s t test. ***p < 0.001. h Control or NLRP4-knockdown Capan-1 cells were treated with DMSO or olaparib (500 nM) for RNA sequencing and were subjected to GO pathway enrichment analysis. i Co-IP experiments were conducted in NLRP4 OE cells, followed by LC‒MS/MS analysis. j Control or NLRP4-knockdown BxPC-3 and Capan-1 cells were treated with the indicated dose of olaparib for 48 h. Western blotting was performed with the indicated antibodies. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pancreatic tumors with higher NLRP4 expression are resistant to olaparib in vivo.a BxPC-3 xenografts obtained from mice in different groups treated with DMSO or olaparib (50 mg/kg per day). b Quantification of the weight of the tumors in different groups treated with DMSO or olaparib (50 mg/kg per day). The analysis of significant differences was performed with Student’s t test. **p < 0.01. c Quantification of tumor volume on the last day in different groups treated with DMSO or olaparib (50 mg/kg per day). The analysis of significant differences was performed with Student’s t test. p < 0.05, ** p <0.01. d Growth curves of cells treated with DMSO or olaparib. The analysis of significant differences was performed with one-way ANOVA. **p < 0.01, ***p < 0.001. e Tumor doubling time in different groups treated with DMSO or olaparib (50 mg/kg per day). The analysis of significant differences was performed with Student’s t test. *p<0.05. f–h Tumors were subjected to immunological staining to detect the indicated marker. f Representative IHC micrographs. Scale bar, 100 μm. g–i The histological score (H score) of the indicated markers was quantified. The analysis of significant differences was performed with Student’s t test. *p < 0.05,**p < 0.01,***p < 0.001. j Western blotting analysis was performed for the indicated antibodies. k Kaplan–Meier survival curves for the specified groups. The p value was calculated by the log rank test. ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

NLRP4 expression promotes pancreatic cancer cell proliferation.a Volcano plot showing the suppression of NLRP4 led to a greater susceptibility to olaparib in pancreatic cancer cell lines. b Immunoblot of NLRP4 in control and NLRP4-knockdown BxPC-3 and Capan-1 cells. BxPC-3 and Capan-1 cells were stably transfected with shNC, shNLRP4-1 and shNLRP4-2, and then MTS assays (c, d) and colony formation assays (e, f) were performed. ***p < 0.001. **p < 0.01. The analysis of significant differences was performed with Student’s t test. g–j Control and NLRP4-knockdown BxPC-3 and Capan-1 cells were injected into the left flank of nude mice. Tumor volumes were measured every 3 days. Tumors were harvested on day 21, photographed and weighed. Tumor weights are shown in (h), BxPC-3 tumor volumes are shown in (i), and Capan-1 tumor volumes are shown in (j). Data are shown as the mean +/- SD (n = 5). ***p < 0.001. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. **p < 0.01. k, l Flow cytometry analysis of annexin V/7-ADD staining in the indicated cell lines. Data are presented as the mean +/- SD from three independent experiments. The analysis of significant differences was performed with Student’s t test. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pancreatic cancer cells with high NLRP4 expression exhibit enhanced levels of autophagy in response to olaparib exposure.a–c Control or NLRP4-knockdown BxPC-3 and Capan-1 cells were treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h. Western blotting analysis and quantification were performed. The analysis of significant differences was performed with Student's t test.**p < 0.01. d–f Effect of NLRP4 on the levels of autophagic flux. The mRFP-GFP-LC3 plasmid was transfected into the indicated cells for 24 h, and then the cells were treated with olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h. Representative images were obtained by laser scanning confocal microscopy. The average fluorescence intensity of autophagic lysosomes (yellow dots in the merged images) and autophagic lysosomes (red in the merged images) in individual cells was quantified. Scale bar, 5 μm. The analysis of significant differences was performed with Student's t test. ***p < 0.001. g–i TEM-based ultrastructure analysis (autophagosomes) in the indicated cells. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

NLRP4 induces mitochondrial ROS generation in pancreatic cancer cells in response to olaparib exposure.a Cells treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h were incubated with an ROS indicator. Representative images were obtained. b, c Cells treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h were incubated with an ROS indicator, and fluorescence intensity was assessed with flow cytometry. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. d, e Cells treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h were incubated with JC-1 working solution, and fluorescence intensity was assessed with flow cytometry. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. f–h Cells treated with olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) plus DMSO or MitoQ for 48 h were incubated with MitoSOX. Representative images were obtained, and the fluorescence intensity was assessed with flow cytometry. The analysis of significant differences was performed with Student’s t test. ***p < 0.001. i Changes in ROS-related genes in control and NLRP4-knockdown Capan-1 cells after olaparib treatment (500 nM olaparib for Capan-1 cells). Scale bar, 5 μm. j Western blotting analysis was performed for the indicated antibodies. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Complementation of NLRP4 rescues DNA repair defects and autophagy levels in NLRP4-knockdown pancreatic cancer cells and results in olaparib resistance.a Representative immunofluorescence micrographs of gamma -H2AX foci in the indicated cells with or without olaparib treatment (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). b TEM-based ultrastructure analysis (autophagosomes) in the indicated cells. c Cells were treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h. Western blotting analysis was performed for the indicated antibodies. d, e Cells treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells) for 48 h were incubated with an ROS indicator, and fluorescence intensity was assessed with flow cytometry. The analysis of significant differences was performed with one-way ANOVA. ***p < 0.001. f, g The indicated cells were treated with DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). MTS assays were performed to investigate the effect of NLRP4 on olaparib resistance. The analysis of significant differences was performed with one-way ANOVA. ***p < 0.001. h Flow cytometry results for annexin V/7-ADD staining in the indicated cell lines after exposure to DMSO or olaparib (5 μM olaparib for BxPC-3 cells and 500 nM olaparib for Capan-1 cells). The analysis of significant differences was performed with Student’s t test. ***p < 0.001. i Schematic model showing that NLRP4 renders pancreatic cancer resistant to PARPi through the promotion of the DNA damage response and ROS-induced autophagy. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41419-024-06984-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Identification of salivary exosomes in periodontitis patients and normal subjects. (A) Transmission electron microscopy of exosomes isolated from human saliva. Scale bar: 100 nm. (B) Exosomes concentration and size distribution by NanoSight analysis of human saliva. (C) Immunoblotting showed the exosomal membrane markers (ALIX, TSG101 CD63, CD9 and CD81), the intracellular protein Calnexin, the marker of autophagosome LC3 and markers of inflammasome (NLRP3 and NLRP4) in exosomes isolated from the saliva of one normal subject (case 01) and one periodontitis patient (case 02). Positive control for Calnexin was TE1 cells, and positive control for LC3 was TE1 cells after starvation for 6 h. Positive control for NLRP3 and NLRP4 was THP-1 cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30923536), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

NALP4 Antibody - BSA Free Summary

• Description: Novus Biologicals Rabbit NALP4 Antibody - BSA Free (NB100-56156) is a polyclonal antibody validated for use in IHC and WB. Anti-NALP4 Antibody: Cited in 6 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: A synthetic peptide corresponding to amino acids 139-157 (FAPKETGKQPRTVIIQGPQ) of human NALP4/PAN2/PYPAF4 was used as immunogen; GenBank no. gi|17064172|gb|AAL35293.1|AF442488_1
• Specificity: NALP4
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NLRP4
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000-1:5000
  • Western Blot 1:1000-1:2000

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Whole antisera
• Preservative: 0.02% Sodium Azide
• Purity: Unpurified

Alternate Names for NALP4 Antibody - BSA Free

• Cancer/testis antigen 58
• CLR19.5
• CT58PYRIN and NACHT-containing protein 2
• FLJ32126
• NACHT, leucine rich repeat and PYD containing 4
• NACHT, LRR and PYD domains-containing protein 4
• NALP4NACHT, LRR and PYD containing protein 4
• NLR family, pyrin domain containing 4
• nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domaincontaining 4
• PAN2Ribonuclease inhibitor 2
• PYPAF4PAAD and NACHT-containing protein 2
• RNH2PYRIN-containing APAF1-like protein 4

Background

NALPs are cytoplasmic proteins that form a subfamily within the larger CATERPILLER protein family. Most short NALPs, such as NALP4, have an N-terminal pyrin (MEFV; MIM 608107) domain (PYD), followed by a NACHT domain, a NACHT-associated domain (NAD), and a C-terminal leucine-rich repeat (LRR) region. The long NALP, NALP1 (MIM 606636), also has a C-terminal extension containing a function to find domain (FIIND) and a caspase recruitment domain (CARD). NALPs are implicated in the activation of proinflammatory caspases (e.g., CASP1; MIM 147678) via their involvement in multiprotein complexes called inflammasomes (Tschopp et al., 2003).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NALP4 Antibody (NB100-56156)(6)

Publications using NB100-56156

• Xiao M, Yang J, Dong M et Al. NLRP4 renders pancreatic cancer resistant to olaparib through promotion of the DNA damage response and ROS-induced autophagy Cell Death Dis 2024-08-26 [PMID: 39187531]
• Wu YL, Pan LH, Yi ZJ et al. c-Myb Dominates TBK1-Mediated Endotoxin Tolerance in Kupffer Cells by Negatively Regulating DTX4 Journal of immunology research 2023-03-31 [PMID: 37032653] (WB, Human)
• Ruscitti P, Berardicurti O, Di Cola I et al. The hyper-expression of NLRP4 characterises the occurrence of macrophage activation syndrome assessing STING pathway in adult onset Still’s disease Clinical and Experimental Immunology 2022-02-02 [PMID: 35467709]
• Yu J, Lin Y, Xiong X et al. Detection of Exosomal PD-L1 RNA in Saliva of Patients With Periodontitis Front Genet. 2019-03-13 [PMID: 30923536] (WB, Human)
• Yamamoto Mayuko, Kondo Eisaku, Takeuchi Makoto et al. miR-155, a Modulator of FOXO3a Protein Expression, Is Underexpressed and Cannot Be Upregulated by Stimulation of HOZOT, a Line of Multifunctional Treg. PLoS One. 2011-01-01 [PMID: 21304824] (WB, Human)
• Jounai N, Kobiyama K, Shiina M et al. NLRP4 negatively regulates autophagic processes through an association with beclin1. J Immunol. 2011-02-01 [PMID: 21209283] (WB)
  
  Details:
  NALP4/PAN2/PYPAF4.1. WB: Endogenous Beclin1 protein was immunoprecipitated from lysed HeLa cells then subject to WB with the NLRP4 (IMG-5743) antibody, Fig 1C.

Bioinformatics

• Gene Symbol: NLRP4
• Entrez:
  • 147945(Human)
  • 243880(Mouse)