Human Apoptosis-associated speck-like protein containing a CARD(PYCARD) ELISA kit

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Human Apoptosis-associated speck-like protein containing a CARD(PYCARD) ELISA kit

Name: Human Apoptosis-associated speck-like protein containing a CARD(PYCARD) ELISA kit
Brand: CUSABIO
SKU: CSB-EL019114HU

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Code: CSB-EL019114HU

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Size:

96T,5×96T,10×96T

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Quantity:

Sample Types: serum, plasma, tissue homogenates

Detect Range: 15.6 pg/mL-1000 pg/mL

Sensitivity: 3.9 pg/mL

Trial Size:

24T ELISA Kit Trial Size (Only USD$150/ kit)
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Target Background

Target Name

PYD and CARD domain containing

Alternative Names

Apoptosis associated speck like protein containing a CARD ELISA Kit; Apoptosis-associated speck-like protein containing a CARD ELISA Kit; ASC ELISA Kit; ASC_HUMAN ELISA Kit; CARD 5 ELISA Kit; CARD5 ELISA Kit; Caspase recruitment domain containing protein 5 ELISA Kit; Caspase recruitment domain protein 5 ELISA Kit; Caspase recruitment domain-containing protein 5 ELISA Kit; hASC ELISA Kit; MGC10332 ELISA Kit; PYCARD ELISA Kit; PYD and CARD domain containing ELISA Kit; PYD and CARD domain containing protein ELISA Kit; PYD and CARD domain-containing protein ELISA Kit; Target of methylation induced silencing 1 ELISA Kit; Target of methylation-induced silencing 1 ELISA Kit; TMS 1 ELISA Kit; TMS ELISA Kit; TMS1 ELISA Kit

Abbreviation

PYCARD

Uniprot NO.

Q9ULZ3

Species

Homo sapiens (Human)

Sample Type

serum, plasma, tissue homogenates

Detection Range

15.6 pg/mL-1000 pg/mL

Sensitivity

3.9 pg/mL

Assay Time

1-5h

Sample Loading Volume

50-100ul

Detection Wavelength

450 nm

Research Area

Cell Biology

Assay Principle

quantitative
The tag type will be determined during production process. If you have specified tag type, please tell us and we will develop the specified tag preferentially.

Measurement

Sandwich

Storage

Store at 2-8°C. Please refer to protocol.

Shelf Life

6 months

Lead Time

3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx.
Note: All of our proteins are default shipped with normal blue ice packs, if you request to ship with dry ice, please communicate with us in advance and extra fees will be charged.

Precision

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of human PYCARD in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
	Sample	Serum(n=4)
1:1	Average %	95
	Range %	90-102
1:2	Average %	90
	Range %	85-99
1:4	Average %	98
	Range %	92-106
1:8	Average %	92
	Range %	86-100

Recovery

The recovery of human PYCARD spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.

Sample Type	Average % Recovery	Range
Serum (n=5)	94	89-98
EDTA plasma (n=4)	95	90-100

Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

pg/ml	OD1	OD2	Average	Corrected
1000	1.834	1.867	1.851	1.713
500	1.517	1.541	1.529	1.391
250	1.213	1.224	1.219	1.081
125	0.892	0.902	0.897	0.759
62.5	0.561	0.578	0.570	0.432
31.2	0.345	0.352	0.349	0.211
15.6	0.256	0.266	0.261	0.123
0	0.137	0.139	0.138

Troubleshooting and FAQs

ELISA kit FAQs

ELISA Data Analysis

ELISA Standard Curve & Curve Expert software

Description

PYCARD, also known as apoptosis-associated speck-like protein containing a CARD, is an adaptor protein that plays a central role in innate immune signaling and inflammasome assembly. This protein contains both a pyrin domain (PYD) and a caspase activation and recruitment domain (CARD), allowing it to bridge interactions between pattern recognition receptors and downstream effector proteins. PYCARD is essential for the formation of inflammasomes, multi-protein complexes that activate inflammatory caspases and process and secrete pro-inflammatory cytokines such as IL-1β and IL-18, making it a key mediator in inflammatory responses and cell death pathways.

This Human PYCARD ELISA kit (CSB-EL019114HU) uses a quantitative sandwich measurement principle to detect PYD and CARD domain containing protein in human samples. The assay works with serum, plasma, and tissue homogenates with a detection range of 15.6 pg/mL to 1000 pg/mL and shows a sensitivity of 3.9 pg/mL. The protocol requires 50-100 μL sample volume and can be completed within 1-5 hours, with detection performed at 450 nm wavelength.

Application Examples

Note: The following application examples are drawn from a selection of publications citing this product. For additional applications, please refer to the full list of references in the "Citations" section.

This ELISA kit has been used in respiratory disease research to quantify ASC protein levels in lung tissue homogenates. The kit supports studies of inflammasome pathway components and their role in inflammatory responses.

• Respiratory disease research: Measuring ASC protein expression in lung tissue to understand inflammasome activation patterns
• Inflammasome pathway studies: Quantifying ASC alongside other inflammasome components including NLRP3 and caspase-1 to evaluate inflammatory signaling cascades
• Cytokine network analysis: Integration with inflammatory mediator profiling to examine relationships between inflammasome components and pro-inflammatory cytokine production
• Tissue-based protein analysis: Application in homogenized tissue samples to measure target protein levels in disease-relevant biological specimens

■ Customer Reviews

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■ Q&A

Can you guarantee that your ELISA can quantify ASC oligomers (ASC specks) in the plasma of patients? Does it recognize other CARD or PYD domain containing proteins?

The kit CSB-EL019114HU is designed for full-length taking a reference of the information from this link http://www.uniprot.org/uniprot/Q9ULZ3 and use the specific raw material.
It detect the general PYCARD.
We get the detection value of serum and plasma is ND--62.5pg/ml.

How do you confirm your kit works? I have cell lines that are
ASC positive and ASC knockout, yet a previous kit I tried from another
company did not work. What positive and negative controls can be used
to confirm this kit really detects ASC?

Thanks for your inquiry!
CSB-EL019114HU
We have made strict screening and quality control from raw materials to finished products to ensure the specificity of our kit and detection effect. This kit is used for quantitative detection of PYCARD. The standard in the kit can be used for validation of the entire reagent system, which can achieve quality control effect. Therefore we no longer provide NC/PC alone. Considering that the customer's sample is cell lines that is ASC positive and ASC knockout, and based on the diversity of samples and the different customers' needs, it is not possible for us to validate all the sample types. So we suggest you do a pre-test if you want to test the cell lysis you mentioned and follow the actual test results.
Pls let me know if you have any further questions. Thank you.

The NLRP3 inhibitor MCC950 inhibits IL-1β production in PBMC from 19 patients with Cryopyrin-Associated Periodic Syndrome and in 2 patients with Schnitzler’s Syndrome SE Corcoran,/,2020
Formation and activity of NLRP3 inflammasome and histopathological changes in the lung of corpses with COVID-19 M Gholaminejhad,/,2022
NLRP3 inflammasome activation and symptom burden in KRAS-mutated CMML patients is reverted by IL-1 blocking therapy L Hurtado-Navarro,/,2023
Systemic inflammation in asymptomatic hyperuricaemia with sonographic crystal deposits, including a comparison with normouricaemia and gout ML Peral-Garrido, S Gómez-Sabater, R Caño,/,2024

Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.

Function(From Uniprot)

Functions as key mediator in apoptosis and inflammation. Promotes caspase-mediated apoptosis involving predominantly caspase-8 and also caspase-9 in a probable cell type-specific manner. Involved in activation of the mitochondrial apoptotic pathway, 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. Involved in macrophage pyroptosis, a caspase-1-dependent inflammatory form of cell death and is the major constituent of the ASC pyroptosome which forms upon potassium depletion and rapidly recruits and activates caspase-1. In innate immune response believed to act as an integral adapter in the assembly of the inflammasome which activates caspase-1 leading to processing and secretion of proinflammatory cytokines. The function as activating adapter in different types of inflammasomes is mediated by the pyrin and CARD domains and their homotypic interactions. Required for recruitment of caspase-1 to inflammasomes containing certain pattern recognition receptors, such as NLRP2, NLRP3, AIM2 and probably IFI16. In the NLRP1 and NLRC4 inflammasomes seems not be required but facilitates the processing of procaspase-1. In cooperation with NOD2 involved in an inflammasome activated by bacterial muramyl dipeptide leading to caspase-1 activation. May be involved in DDX58-triggered proinflammatory responses and inflammasome activation. Isoform 2 may have a regulating effect on the function as inflammasome adapter. Isoform 3 seems to inhibit inflammasome-mediated maturation of interleukin-1 beta. In collaboration with AIM2 which detects cytosolic double-stranded DNA may also be involved in a caspase-1-independent cell death that involves caspase-8. In adaptive immunity may be involved in maturation of dendritic cells to stimulate T-cell immunity and in cytoskeletal rearrangements coupled to chemotaxis and antigen uptake may be involved in post-transcriptional regulation of the guanine nucleotide exchange factor DOCK2; the latter function is proposed to involve the nuclear form. Also involved in transcriptional activation of cytokines and chemokines independent of the inflammasome; this function may involve AP-1, NF-kappa-B, MAPK and caspase-8 signaling pathways. For regulation of NF-kappa-B activating and inhibiting functions have been reported. Modulates NF-kappa-B induction at the level of the IKK complex by inhibiting kinase activity of CHUK and IKBK. Proposed to compete with RIPK2 for association with CASP1 thereby down-regulating CASP1-mediated RIPK2-dependent NF-kappa-B activation and activating interleukin-1 beta processing. Modulates host resistance to DNA virus infection, probably by inducing the cleavage of and inactivating CGAS in presence of cytoplasmic double-stranded DNA.

Gene References into Functions

Cryo-EM structures of ASC and NLRC4 CARD filaments reveal a unified mechanism of nucleation and activation of caspase-1. PMID:30279182
ASC specks as a putative biomarker of pyroptosis in myelodysplastic syndromes PMID:30072146
results suggest that ASC, as a negative regulator of the MAVS-mediated innate immunity, may play an important role in host protection upon virus infection PMID:29280086
PYCARD gene and its transcript variant may play a critical and regulative role in the inflflammatory response of primary gout patients with different phases and Chinese medicine syndromes. PMID:29086221
ASC may be involved in tumor suppression and cell death via Bcl-2 and phosphor Src. PMID:29459573
Data show that in HK-2 cells and unilateral nephrectomy model, ASC expression level is significantly augmented after treatment with contrast media. Its silencing attenuates contrast-induced apoptosis in HK-2 cell. PMID:27721494
ASC specks released by microglia bind to amyloid-beta and increase amyloid-beta oligomer and aggregate formation, acting as an inflammation-driven cross-seed for amyloid-beta pathology PMID:29293211
ASC contributes to oral cavity squamous cell carcinoma metastasis, and high-level ASC expression is a marker for poor prognosis in OSCC patients PMID:27367024
ASC CpG methylation may prove to be a primary regulator of the pathogenesis of chronic inflammatory diseases such as heart failure. PMID:26700661
besides its role in the inhibition of the NF-kappaB pathway, NLRC3 interferes with the assembly and activity of the NALP3 inflammasome complex by competing with ASC for pro-caspase-1 binding PMID:28584053
ASC Induces Apoptosis via Activation of Caspase-9 by Enhancing Gap Junction-Mediated Intercellular Communication.( PMID:28056049
These data revealed that cross-linking of ASC(PYD) filaments via ASC(CARD) mediates the assembly of ASC foci. PMID:27069117
Down-regulation of mRNA expression was found in cases in which CASP8, TMS1 and DAPK were hypermethylated. PMID:28361856
loss of ASC driven tumor development is counterbalanced in the identical cell by the inhibition of pro-tumorigenic inflammation in the tumor cell itself PMID:27768771
the deubiquitinating enzyme USP50 binds to the ASC protein and subsequently regulates the inflammasome signaling pathway. PMID:28094437
ASC self-associates and binds NLRP3 PYD through equivalent protein regions, with higher binding affinity for the latter. These regions are located at opposite sides of the protein allowing multimeric complex formation previously shown in ASC PYD fibril assemblies. PMID:27432880
Our data identify RIPK3 and the ASC inflammasome as key tumor suppressors in AML. PMID:27411587
The role of the danger signals ASC and HMGB1 in the Fusobacterium nucleatum infection of gingival epithelial cells. PMID:26687842
Data show that NOD-like receptor signaling genes NOD2, PYCARD, CARD6, and IFI16 are upregulated in psoriatic epidermis. PMID:26976200
The methylated status of ASC/TMS1 promoter had the potential applicability for clinical evaluation the prognosis of gastric cancer PMID:26260914
it appears that ASC transcript expression may be a surrogate marker for depression and may represent a link between depression and the altered immune responses observed in these categories of individuals with elevated depressive symptoms. PMID:26750863
The proteins of NLRP3, ASC, and caspase-1 were observed in infiltrating inflammatory cells in cholesteatoma and chronic otitis media. PMID:26457439
ASC/TMS1 methylation was significantly correlated with higher tumor nuclear grade. ASC/TMS1 is a novel functional tumor suppressor in renal carcinogenesis. PMID:26093088
ASC Induces Procaspase-8 Death Effector Domain Filaments PMID:26468282
ASC interacts with NALP3 and caspase-1 via different domains. PMID:25567507
mRNA levels of Apoptosis-associated Speck-like protein were significantly higher in freshly isolated PBMCs from Chronic recurrent multifocal osteomyelitis patients in active disease than in healthy controls. PMID:25061439
The proteins (HSP90b, TSM1 and L-plastin) in the current study may hold potential in differentiating between melanoma and benign nevi in diagnostically challenging cases. PMID:25191796
caspase-1/ASC inflammasomes play a significant role in the activation of IL-1beta/ROS and NF-kappaB signaling of cytokine gene expression for T. cruzi control in human and mouse macrophages. PMID:25372293
Neutralization of ASC improves sperm motility in men with spinal cord injuries. PMID:25205754
Transcriptome analysis of human adipocytes implicates the NOD-like receptor pathway (NLRP3, PYCARD) in obesity-induced adipose inflammation. PMID:25011057
Data indicate that apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is highly expressed in medulloblastomas. PMID:24469054
R42W mutation had a significant effect on structure and increased stability. Although the R42W mutant exhibited reduced interaction with ASC PMID:25006247
ASC PYD prevented complex formation with NALP3 PYD in vitro PMID:24585381
Identify a novel innate immune signaling pathway (NLRP3-ASC-caspase-1-IL-1beta) activated by Ni(2+). PMID:24158569
this study was to investigate the mRNA levels of AIM2 and ASC in a lymphocyte cell line (Jurkat) before and after MiR-143 introducing. PMID:23811549
this study reports an interaction between promyelocytic leukemia protein and apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC). PMID:24407287
PYCARD methylation is associated with colon cancer. PMID:24169962
Activated AIM2 and NLRP3 nucleate PYD filaments of ASC, which, in turn, cluster the CARD of ASC. ASC thus nucleates CARD filaments of caspase-1, leading to proximity-induced activation.studies revealed a universal assembly process for ASC-dependent inflammasomes in both ALR and NLR families that involves nucleation-induced polymerization. PMID:24630722
Study shows that T. gondii-induced IL-1beta production is dependent on the classical inflammasome components caspase-1 and ASC.Additionally, a role for a specific parasite factor, dense granule protein GRA15, in T. gondii induction of IL-1beta was demonstrated. PMID:23839215
Reactivation of ASC protein expression in LS174T cells promotes sodium butyrate induced apoptosis. PMID:23064206
The findings of this work may suggest a crucial relationship between mutant MEFV/pyrin and remarkable ASC up-regulation in familial Mediterranean fever inflammation. PMID:22934972
These findings suggest stage-dependent dual roles of ASC in melanoma tumorigenesis. PMID:22931929
central role of CARDs in the formation of ASC signalling platforms PMID:23110696
ASC PYD can simultaneously self-associate and interact with NLRP3, rationalizing the model whereby ASC self-association upon recruitment to NLRP3 promotes clustering and activation of procaspase-1. PMID:23066025
ASC in different tissues may influence tumor growth in opposite directions. PMID:23090995
The study conclude that the frequency of TMS1/ASC gene methylation in cervical cancer is rare and have no any critical role in development of cervical cancer. PMID:19258216
Gene expression profiles of ASC or CatB deficient human DCs show marked overlap with downregulation of genes implicated in DC function. PMID:22732093
the requirement of TLR2/MyD88/NF-kappaB pathway (first signal) and ROS/potassium efflux (second signal) for NLRP3/ASC inflammasome formation, leading to caspase-1 activation and subsequent IL-1beta release during RSV infection PMID:22295065
Hypermethylation of ASC is associated with cholangiocarcinoma. PMID:22230750
Caspase-1 protein induces apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC)-mediated necrosis independently of its catalytic activity. PMID:21832064

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Subcellular Location

Cytoplasm. Inflammasome. Endoplasmic reticulum. Mitochondrion. Nucleus.; Golgi apparatus membrane.

Tissue Specificity

Widely expressed at low levels. Detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, colon and at lower levels in placenta, liver and kidney. Very low expression in skeletal muscle, heart and brain. Expressed in lung epithelial

Database Links

HGNC: 16608

UNIGENE: Hs.499094

KEGG: hsa:29108

STRING: 9606.ENSP00000247470

OMIM: 606838

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