Anti-ASC/PYCARD Antibody, clone 1C19 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1756 - 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. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in germinal centers & interfollicular area of human tonsil tissue sections.
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. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in germinal centers & interfollicular area of human tonsil tissue sections.
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. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in germinal centers & interfollicular area of human tonsil tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cells was performed using a 1:100 dilution of Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cells was performed using a 1:100 dilution of Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of THP-1 cells was performed using a 1:100 dilution of Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and the cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology THP-1 (Lane 1) and HL-60 (Lane 2) cell lysates were probed with Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates ASC/PYCARD (~22 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or ASC (PYCARD) peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal. The binding of this antibody to ASC (PYCARD) peptide displayed a KD of 1.1 x 10-8 whereas binding to non-specific control peptide displayed a KD of 1.3 x 10-4.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million THP-1 cells was performed using 1 μg of Cat. No. ZRB1756, Anti-ASC/PYCARD, clone 1C19 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1C19, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | C-terminal half |
| 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 21.63 kDa |
| observed mol wt ~22 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_037390  |
| purified by | using Protein A |
| 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 |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q9ULZ3 |
| Application: | Quality Control Testing Evaluated by Western Blotting in THP-1 cell lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected ASC/PYCARD in THP-1 cell lysate. Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected ASC/PYCARD in HL-60 cell lysate. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected ASC/PYCARD in THP-1 cells. Flow Cytometry Analysis: 1 μg from a representative lot detected ASC/PYCARD in one million THP-1 cells. Affinity Binding Assay: A representative lot of this antibody bound ASC with a KD of 1.1 x 10-8 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected ASC/PYCARD in human tonsil tissue sections. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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 22 amino acids from the C-terminal half of human Apoptosis-associated speck-like protein containing a CARD (ASC/PYCARD). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. 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 1C19 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Apoptosis-associated speck-like protein containing a CARD (ASC/PYCARD). It targets an epitope within 22 amino acids from the C-terminal half. |
| 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: | Apoptosis-associated speck-like protein containing a CARD (UniProt: Q9ULZ3; also known as hASC, Caspase recruitment domain-containing protein 5, PYD and CARD domain-containing protein, Target of methylation-induced silencing 1) is encoded by the PYCARD (also known as ASC, CARD5, TMS1) gene (Gene ID: 29108) in human. PYCARD is widely expressed at low levels and is detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, and colon. At significantly reduced levels it is also found in placenta and liver, and kidney. It is detected in the nucleus, mitochondria, endoplasmic reticulum, cytoplasm, and Golgi apparatus. It is localized primarily to the nucleus in resting monocytes/macrophages and rapidly redistributed to the cytoplasm upon pathogen infection. PYCARD functions as key mediator in apoptosis and inflammation and promotes caspase-mediated apoptosis. It is involved in activation of the mitochondrial apoptotic pathway where it promotes caspase-8-dependent proteolytic maturation of BID independently of FADD in certain cell types and also mediates mitochondrial translocation of Bax and activates Bax-dependent apoptosis coupled to activation of caspase-9, -2 and -3. In innate immune response, it acts as an integral adapter in the assembly of the inflammasome that activates caspase-1 leading to processing and secretion of proinflammatory cytokines. Its function as an activating adapter in different types of inflammasomes is mediated by the pyrin (aa 1-91) and CARD (aa 107-195) domains and their homotypic interactions. It is shown to be essential for recruitment of caspase-1 to inflammasomes containing certain pattern recognition receptors, such as NLRP2, NLRP3, AIM2 and probably IFI16. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Bryan, NB., et al. (2009). J. Immunol. 182(5); 3173-3182; Ohtsuka, T., et al. (2004). Nat. Cell Biol. 6(2); 121-128). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |