Anti-NLRP3 Antibody, clone 4B4 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1762 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in subsets of non-germinal center cells of human tonsil tissue.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in subsets of non-germinal center cells of human tonsil tissue.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in subsets of non-germinal center cells of human tonsil tissue.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 (A&B) cell line and THP-1 treated (C&D) with 200 nM PMA for 24 hours and 10 μg/mL LPS for 3 hours were performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 (A&B) cell line and THP-1 treated (C&D) with 200 nM PMA for 24 hours and 10 μg/mL LPS for 3 hours were performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 (A&B) cell line and THP-1 treated (C&D) with 200 nM PMA for 24 hours and 10 μg/mL LPS for 3 hours were performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 (A&B) cell line and THP-1 treated (C&D) with 200 nM PMA for 24 hours and 10 μg/mL LPS for 3 hours were performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 (A&B) cell line and THP-1 treated (C&D) with 200 nM PMA for 24 hours and 10 μg/mL LPS for 3 hours were performed using a 1:100 dilution of Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Human Spleen tissue lysate was probed with Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates NLRP3 (~95 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of NLRP3 peptide (Run 1) or non-specific control peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1762, Anti-NLRP3, clone 4B4 ZooMAb® Rabbit Monoclonal. The binding of this antibody to NLRP3 peptide displayed a KD of 1.9 x 10-6, whereas binding to non-specific control peptide displayed a KD of 2.0 x 10-3.
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Chemical products should be designed to affect their desired function while minimizing their toxicity.
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| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 4B4, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt ~118.17 kDa |
| observed mol wt ~95 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q96P20  |
| Application: | Quality Control Testing Evaluated by Western Blotting in Human Spleen tissue lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected NLRP3 in Human Spleen tissue lysate.Tested Applications Affinity Binding Assay: A representative lot of this antibody bound NLRP3 peptides with a KD of 1.9 x 10-6 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected NLRP3 in human tonsil tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected NLRP3 in THP-1 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-NLRP3, clone 4B4 ZooMAb®, Cat. No. ZRB1762, is a recombinant Rabbit monoclonal antibody that specifically targets NLRP3 and is tested for use in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 15 amino acids from the N-terminal domain of human NLRP3. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 4B4 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects human NLRP3 protein. It targets an epitope within 15 amino acids from the N-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | NACHT, LRR and PYD domains-containing protein 3 (UniProt: Q96P20; also known as Angiotensin/vasopressin receptor AII/AVP-like, Caterpiller protein 1.1, CLR1.1, Cold-induced autoinflammatory syndrome 1 protein, Cryopyrin, PYRIN-containing APAF1-like protein 1, NLRP3) is encoded by the NLRP3 (also known as C1orf7, CIAS1, NALP3, PYPAF1) gene (Gene ID: 114548) in human. NLRP3 serves as the sensor component of the NLRP3 inflammasome and plays a crucial role in innate immunity and inflammation. It is predominantly expressed in macrophages and its expression is also observed in dendritic cells, B- and T-cells, and in lung epithelial cells. NLRP3 can be induced by activators of Toll-like receptors, bacterial lipopolysaccharides, and by TNF. In response to pathogens and other damage-associated signals, it initiates the formation of the inflammasome polymeric complex that is composed of NLRP3, PYCARD and CASP1. Recruitment of proCASP1 to the inflammasome is shown to promote its activation and CASP1-catalyzed IL-1B and IL-18 maturation and secretion in the extracellular milieu. Under resting conditions, NLRP3 is autoinhibited and it is activated in response to several signals, including extracellular ATP, reactive oxygen species, K+ efflux, crystals of monosodium urate or cholesterol, and amyloid-beta fibers. Independently of inflammasome activation, it is reported to regulate the differentiation of T helper 2 (Th2) cells and is shown to play a role in Th2 cell-dependent asthma and tumor growth. NLRP3 undergoes polyubiquitination at lysine 48 and 63. However, this ubiquitination does not lead to its degradation, but inhibits inflammasome activation. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |