Human Caspase 1(Casp-1) ELISA Kit

Instructions
Code CSB-E13025h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Caspase 1 (Casp-1) ELISA Kit is suitable for quantitative detection of Casp-1 in serum, plasma, and tissue homogenates. Samples or standards are added into the microwells and bind to the anti-human Casp-1 antibody immobilized on the microplate. A sandwich is formed by the addition of the biotin-conjugated Casp-1 antibody. The TMB substrate solution is added that reacts with the enzyme-antibody-target complex to elicit a chromogenic reaction. The intensity of the solution color is directly proportional to the concentration of Casp-1 present in the original specimen. This kit has been verified with a wide detection range (12.5 pg/mL-800 pg/mL), high sensitivity, excellent specificity, precision less than 10%, good linearity, high recovery, and good consistency between lots. Refer to the product instructions for more verification details.

Casp-1 is synthesized as an inactive zymogen that is recruited and activated by inflammasomes. It plays a crucial role in cell immunity as an inflammatory response inhibitor. After activation by inflammasomes, Casp-1 initiates a pro-inflammatory response through the cleavage and subsequent activation of the two inflammatory cytokines, IL-1β and IL-18 as well as gasdermin D (GSDMD)-mediated pyroptosis, thus killing the infectious agent. In the absence of GSDMD, Casp-1 induces apoptosis involves the Bid-caspase-9-caspase-3 axis, which can elicit secondary necrosis/pyroptosis mediated by GSDME/DFNA5.

Target Name caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase)
Alternative Names CASP-1 ELISA Kit; CASP1 ELISA Kit; CASP1_HUMAN ELISA Kit; Caspase 1 ELISA Kit; Caspase-1 subunit p10 ELISA Kit; ICE ELISA Kit; IL-1 beta-converting enzyme ELISA Kit; IL-1BC ELISA Kit; IL1 beta converting enzyme ELISA Kit; IL1B convertase ELISA Kit; Interleukin 1 beta convertase ELISA Kit; Interleukin 1B converting enzyme ELISA Kit; Interleukin-1 beta convertase ELISA Kit; Interleukin-1 beta-converting enzyme ELISA Kit; p45 ELISA Kit
Abbreviation CASP1
Uniprot No. P29466
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 12.5 pg/mL-800 pg/mL
Sensitivity 3.1 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cell Biology
Assay Principle quantitative
Measurement Sandwich
Precision
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.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human Casp-1 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 % 86  
Range % 81-92  
1:2 Average % 101  
Range % 96-105  
1:4 Average % 94  
Range % 89-98  
1:8 Average % 103  
Range % 97-108  
Recovery
The recovery of human Casp-1 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) 101 97-105  
EDTA plasma (n=4) 92 88-96  
             
             
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  
800 2.112 1.984 2.048 1.920  
400 1.634 1.537 1.586 1.458  
200 1.124 1.085 1.105 0.977  
100 0.718 0.693 0.706 0.578  
50 0.389 0.394 0.392 0.264  
25 0.262 0.274 0.268 0.140  
12.5 0.189 0.182 0.186 0.058  
0 0.128 0.127 0.128    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human Casp-1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled Casp-1 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Data

Function Thiol protease that cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes. Important for defense against pathogens. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Can also promote apoptosis. Upon inflammasome activation, during DNA virus infection but not RNA virus challenge, controls antiviral immunity through the cleavage of CGAS, rendering it inactive
Gene References into Functions
  1. ABT-737 elicited an anti-atopic dermatitis (AD) activity via suppression of caspase-1 activation in AD in vitro and in vivo models. Therefore, this study provides important information regarding the use of anticancer drugs for controlling allergic inflammatory diseases. PMID: 29957081
  2. Cryo-EM structures of ASC and NLRC4 CARD filaments reveal a unified mechanism of nucleation and activation of caspase-1 PMID: 30279182
  3. The increased expression of NLRP3 and caspase-1 in fetal membrane and placental tissues may be associated with the development of premature rupture of membrane. PMID: 29545514
  4. inflammasome activation and pyroptosis in human microglia and ODCs in vitro after exposure to inflammatory stimuli and demonstrate caspase-1 inhibition by the small-molecule inhibitor VX-765 in both cell types. GSDMD inhibition by siRNA transduction suppressed pyroptosis in human microglia. PMID: 29895691
  5. Increased production of caspase-1 in the gut-associated lymphoid tissue and peripheral blood of HIV infected patients. PMID: 29672590
  6. Low CASP1 expression is associated with prostate cancer. PMID: 28388569
  7. These findings confirm the involvement of caspase-1 in non-classical secretion mechanisms and open novel perspectives for the extracellular function of secreted GBP-1. PMID: 28272793
  8. this study demonstrates that the G+7/in6A and A10370-G polymorphisms of the CASP1 gene are associated with increased risk of developing acute coronary syndrome in Mexican population PMID: 28456882
  9. After genotype calling and quality control filtering with exclusion of 3 cases and 3 controls, association analysis was performed across 76 directly genotyped SNPs in NLRP1, CARD, and CASP1 genes, adjusting for age, sex, and population stratification PMID: 29438387
  10. Caspase-1 role in pyroptosis in glioma cells.Mir-214 regulates caspase-1 expression in glioma. PMID: 28244850
  11. Data show that cyclic stretch activated the nucleotide-binding oligomerization domain-like receptor containing pyrin domain 1 and 3 (NLRP1 and NLRP3) inflammasomes and induced the release of IL-1beta and pyroptosis via a caspase-1-related mechanism in human periodontal ligament cells (HPDLCs). PMID: 27626170
  12. Caspase-1 directly cleaves alpha-Synuclein, generating a highly aggregation-prone species in neurons. PMID: 27482083
  13. the activation of Rho GTPases by the CNF1 toxin during E. coli-triggered bacteremia leads to a GR1(+)cell-mediated efficient bacterial clearing and improves host survival. Host alarm requires the Caspase-1/IL-1beta signaling axis. PMID: 26492464
  14. Nodakenin inhibited the mRNA expression and production of pro-inflammatory cytokines and caspase-1 activation PMID: 28407357
  15. sickle red blood cell induced TLR9, NLRP3, Caspase-1, IL-1beta and IL-18 expression and induced IL-1beta, LTB4 and nitrite production in PBMC cultures. PMID: 27045344
  16. Data show that the serine protease inhibitor B9 (serpinB9) mediated caspase-1 inhibition regulates IL-1beta release in monocytes. PMID: 26992230
  17. study uncovers a novel mechanism of G9A promoting tumor cell growth and invasion by silencing CASP1, and implies that G9A may serve as a therapeutic target in treating Non-small-cell lung cancer. PMID: 28383547
  18. Caspase-1 polymorphisms may play a role in Chagas cardiomyopathy development and could serve as markers to identify individuals at higher risk for priority treatment. PMID: 28954073
  19. 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
  20. Data, including data from studies using recombinant fusion forms of GSDMD (gasdermin D), suggest that GSDMD participates in inflammasome-dependent pyroptosis of macrophages in response to various stimuli; this mechanism involves proteolysis of GSDMD by caspase-1 and caspase-11. PMID: 28726636
  21. Proteases caspase-1 and caspase-8 have redundant roles in cleaving IL-1beta and promoting osteomyelitis. [review] PMID: 27148834
  22. Patients with NLRP1-associated autoinflammation with arthritis and dyskeratosis syndrome had increased systemic CASP1. PMID: 27965258
  23. The biochemical function of NLPR3 inflammasomes is to activate caspase-1, which leads to the maturation of interleukin 1 beta and IL-18 and the induction of pyroptosis, a form of cell death. (Review) PMID: 27669650
  24. The structure of the human caspase-1 CARD domain (caspase-1(CARD)) filament solved by cryo-electron microscopy. PMID: 27043298
  25. Caspase-1-mediated downregulation of PPARgamma was important in the late stage of monocyte-macrophage differentiation; however, PPARgamma protein levels had little effect on the early stage differentiation. PMID: 28052562
  26. Inflammasome NLRP3-caspase-1-mediated degradation of smooth muscle cell contractile proteins may contribute to aortic biomechanical dysfunction and aortic aneurysm/dissection development. PMID: 28153878
  27. Study demonstrates that the expression of the NLRP3-caspase-1-IL-18 axis is highly expressed in the peripheral blood mononuclear cells of patients with bullous pemphigoid (BP), and correlated with disease activity, suggesting its involvement in the pathogenesis and progression of BP. PMID: 27174093
  28. High expression of CASP1 is associated with osteosarcoma. PMID: 28000894
  29. This study demonstrated that dysregulated expression of NLRP3-caspase-1-IL-1beta axis was observed in patients with MM, suggesting they might be involved in the pathogenesis of MM. PMID: 26146985
  30. The NLRP3/caspase-1/IL-1beta axis is active in cartilaginous endplates of patients with Modic changes and inflammatory cascades can exacerbate the cartilaginous endplate degeneration which may act as a trigger for intervertebral disc degeneration and low back pain. PMID: 27146654
  31. High expression of NLRP3, NLRC4, and CASP1 in background non-tumorous liver is significantly correlated with poor prognosis of patients after resection of hepatocellular carcinoma. PMID: 28011505
  32. The proteins of NLRP3, ASC, and caspase-1 were observed in infiltrating inflammatory cells in cholesteatoma and chronic otitis media. PMID: 26457439
  33. The results indicate that acteoside may act as a regulator of the IL-32 induced immune responses. PMID: 26453510
  34. These results suggest that released caspase-1 exists in a unique complex that is functionally stable and protected from immunodepletion whereas cell-extract generated active caspase-1 is rapidly inhibited in the cytosolic milieu. PMID: 26599267
  35. Genetic polymorphisms in Nalp3 and caspase-1 may be associated with individual susceptibility to silicosis, especially when the polymorphisms interact with age, cumulative dust exposure, or smoking status. PMID: 26496436
  36. In contrast, inhibition of the protease activity of Kgp, RgpA or RgpB increased the human caspase-1-activating potential of wild-type Porphyromonas gingivalis, indicating an inhibitory effect of the collaborative action of gingipains. PMID: 25759090
  37. NLRP3 and caspase-1 are expressed in the odontoblast layers in normal human dental pulp tissue, whereas in inflamed pulp tissue, the odontoblast layers are disrupted and dental pulp cells are positive for NLRP3 and caspase-1. PMID: 25684031
  38. the rs713875 IBD risk polymorphism increases MTMR3 expression, which modulates pattern recognition receptor (PRR)-induced outcomes, including increased induced caspase-1 activation PMID: 26240347
  39. specific caspase cleavage sites in Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen function to blunt apoptosis as well as interfere with the caspase-1-mediated inflammasome PMID: 26218605
  40. three cofactor independent human monoclonal aPL can induce transcription of NLRP3 and caspase-1 resulting in inflammasome activation specific for NLRP3, which depends fully on activation of endosomal NADPH-oxidase-2 (NOX2) by aPL PMID: 25589411
  41. Internalized Cryptococcus neoformans Activates the Canonical Caspase-1 and the Noncanonical Caspase-8 Inflammasomes. PMID: 26466953
  42. In patients with HIV, serum CASP1 levels in a group with high CD4 cell counts increased rapidly then decreased within a short time during early HIV-1 infection. In contrast, CASP1 levels in the CD4Low group were increased after 1 year of HIV-1 infection. PMID: 25806508
  43. production of 20-kDa IL-1beta may act to limitIL-1beta signaling by reducing the pool of pro-IL-1beta availability for caspase-1 cleavage PMID: 26324708
  44. The study shows that A1AT does not inhibit human monocyte caspase-1. PMID: 25658455
  45. Data suggest activation of CASP1 (caspase-1) and secretion of IL18 (interleukin 18)/IFNG (interferon-gamma) in mucosal explants of ileum/colon are related to intensity of inflammatory lesions in patients with active, long-standing Crohn's disease. PMID: 26168332
  46. Interleukin-1alpha Activity in Necrotic Endothelial Cells Is Controlled by Caspase-1 Cleavage of Interleukin-1 Receptor-2 PMID: 26324711
  47. the inflammatory CASP1 is linked to several genes involved in cholesterol homeostasis. These correlations resulted in altered GBM (Glioblastoma Multiforme), in particular for CASP1 PMID: 25330190
  48. caspase 1 was higher in degenerated intervertebral discs than in healthy controls. PMID: 25284686
  49. ASC interacts with NALP3 and caspase-1 via different domains. PMID: 25567507
  50. Changes in the tertiary structure of mutated pyrin B30.2 domain interrupting the formation of the pyrin-caspase-1 complex are related to the manifestations of Mediterranean familial fever. PMID: 26510601

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Subcellular Location Cytoplasm
Protein Families Peptidase C14A family
Tissue Specificity Expressed in larger amounts in spleen and lung. Detected in liver, heart, small intestine, colon, thymus, prostate, skeletal muscle, peripheral blood leukocytes, kidney and testis. No expression in the brain.
Database Links

HGNC: 1499

OMIM: 147678

KEGG: hsa:834

STRING: 9606.ENSP00000410076

UniGene: Hs.2490

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