Caspase-1 Antibody Pair [Biotin]
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-78858
Key Product Details
Assay Type
Sandwich ELISA
Assay Range
0.15ng/ml - 10 ng/ml
Sample Type
Cell Culture Supernatant (100 ul)
Sensitivity
50 pg/ml
Reactivity
Mouse
Product Specifications
Description
Sandwich ELISA for the quantitative determination of Mouse Caspase-1
Sample Volume Required
100 ul
Conjugate
Biotin
Application Notes
1. Dilute the desired amount of Coating Antibody (COAT) (1 mg/ml) (mouse monoclonal antibody) to 2 ug/ml in PBS without carrier protein and use it fresh.
2. Dilute the desired amount of Detection Antibody (DET) (0.5 mg/ml) (biotinylated mouse monoclonal antibody) to 1 ug/ml in ELISA Buffer and use it fresh.
3. Reconstitute the Streptavidin-HRP (STREP) with 100 ul ELISA Buffer. After reconstitution, prepare aliquots and store the reconstituted STREP at -20C. Avoid freeze/thaw cycles. Dilute the reconstituted Streptavidin-HRP to the working concentration by adding 10 ul in 10 ml of ELISA Buffer (1:1000).
4. Reconstitute the Standard Protein (STD) (recombinant mouse Caspase-1) with 100 ul ELISA Buffer to obtain a concentration of 10 ug/ml. After reconstitution, prepare aliquots and store the reconstituted standard at -20C. Avoid freeze/ thaw cycles. A standard curve using 2-fold serial dilutions in ELISA Buffer is recommended. Suggested standard points are 10 ng/ml, 5 ng/ml, 2.5 ng/ml, 1.25 ng/ml, 0.625 ng/ml, 0.3125 ng/ml, 0.15625 ng/ml and 0 ng/ml.
2. Dilute the desired amount of Detection Antibody (DET) (0.5 mg/ml) (biotinylated mouse monoclonal antibody) to 1 ug/ml in ELISA Buffer and use it fresh.
3. Reconstitute the Streptavidin-HRP (STREP) with 100 ul ELISA Buffer. After reconstitution, prepare aliquots and store the reconstituted STREP at -20C. Avoid freeze/thaw cycles. Dilute the reconstituted Streptavidin-HRP to the working concentration by adding 10 ul in 10 ml of ELISA Buffer (1:1000).
4. Reconstitute the Standard Protein (STD) (recombinant mouse Caspase-1) with 100 ul ELISA Buffer to obtain a concentration of 10 ug/ml. After reconstitution, prepare aliquots and store the reconstituted standard at -20C. Avoid freeze/ thaw cycles. A standard curve using 2-fold serial dilutions in ELISA Buffer is recommended. Suggested standard points are 10 ng/ml, 5 ng/ml, 2.5 ng/ml, 1.25 ng/ml, 0.625 ng/ml, 0.3125 ng/ml, 0.15625 ng/ml and 0 ng/ml.
Kit Contents for Caspase-1 Antibody Pair [Biotin]
- 1 vial coating antibody (120 ug) [COAT]
- 1 vial detection antibody (60 ug) [DET]
- 1 vial standard protein (1 ug) (lyophilized) [STD]
- 1 vial streptavidin-HRP (10 ug) (lyophilized) [STREP]
Preparation and Storage
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: Caspase-1
Given the role of IL-1beta in inflammation, it makes sense that many diseases and pathologies have been associated with dysregulation of caspase-1 activation and the inflammasome (3, 4). The inflammasome is a multiprotein complex comprised of Nod-like receptor (NLR) family members and the adapter ASC (apoptosis-associated speck-like protein containing a CARD) which are crucial for capase-1 activation (3-5). For instance, the neuronal apoptosis inhibitory protein (NAIP)/NLRC4 inflammasome has been associated with colorectal cancer, breast cancer, and glioma pathogenesis (5). Caspase-1 activation and mutations in the inflammasome have also been linked to Chron's disease and Alzheimer's disease (4). In addition to immune and inflammatory related disorder, the inflammasome has been linked to metabolic and obesity related disorders including diabetes and cardiovascular disease (6). Finally, caspase-1 deficient mice exhibit enhanced epithelial cell proliferation in the colon, increased tumor formation, and reduced apoptosis (1). A more thorough understanding of the inflammasome-caspase-1 signaling pathway will be important for understanding disease pathology and potential therapeutic development.
Alternative names for caspase-1 includes CASP1, CASP1 nirs variant 1, EC 3.4.22.36, ICE, IL-1 beta-converting enzyme, IL1BC, IL1BCE, IL1B-converstase, interleukin-1 beta convertase, and p45.
References
1. Shalini, S., Dorstyn, L., Dawar, S., & Kumar, S. (2015). Old, new and emerging functions of caspases. Cell death and differentiation. https://doi.org/10.1038/cdd.2014.216
2. Chang, H. Y., & Yang, X. (2000). Proteases for cell suicide: functions and regulation of caspases. Microbiology and molecular biology reviews: MMBR. https://doi.org/10.1128/mmbr.64.4.821-846.2000
3. Vanaja, S. K., Rathinam, V. A., & Fitzgerald, K. A. (2015). Mechanisms of inflammasome activation: recent advances and novel insights. Trends in cell biology. https://doi.org/10.1016/j.tcb.2014.12.009
4. Franchi, L., Eigenbrod, T., Munoz-Planillo, R., & Nunez, G. (2009). The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis. Nature immunology. https://doi.org/10.1038/ni.1703
5. Kay, C., Wang, R., Kirkby, M., & Man, S. M. (2020). Molecular mechanisms activating the NAIP-NLRC4 inflammasome: Implications in infectious disease, autoinflammation, and cancer. Immunological reviews. https://doi.org/10.1111/imr.12906
6. Pham, D., Park, P. (2020). Recent insights on modulation of inflammasomes by adipokines: a critical event for the pathogenesis of obesity and metabolism-associated diseases. Archives of Pharmacal Research. https://doi.org/10.1007/s12272-020-01274-7
Alternate Names
CASP1, Caspase1, ICE
Gene Symbol
CASP1
Additional Caspase-1 Products
Product Documents for Caspase-1 Antibody Pair [Biotin]
Product Specific Notices for Caspase-1 Antibody Pair [Biotin]
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Pairs are guaranteed for 6 months from date of receipt.
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