Caspase-1 Overexpression Lysate

Name: Caspase-1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-05896

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-05896

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Caspase-1 Overexpression Lysate

Caspase-1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC218982

Accession#: NM_033294

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

29.6 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for Caspase-1 Overexpression Lysate

Western Blot: Caspase-1 Overexpression Lysate [NBP2-05896]

Western Blot: Caspase 1 Overexpression Lysate (Adult Normal) [NBP2-05896] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Caspase 1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Caspase-1

Caspase-1, also known as interleukin-1-beta (IL-1beta) -converting enzyme (ICE), is a member of the cysteine-aspartic protease (caspase) family (1). The caspase family are aspartic proteolytic enzymes that play a key role in apoptotic cell death and inflammation (1). Caspase-1 is synthesized as an inactive proenzyme with a theoretical molecular weight of approximately 45 kDa and is 404 amino acids (aa) in length. The caspase-1 protein structure contains an N-terminal CARD (caspase recruitment domain), a large 20kDa P20 subunit (aa 120-297), and a small 10kDa P10 subunit (aa 317-404) (1, 2). The proenzyme undergoes cleavage at aspartic acid (Asp) residues to produce the active caspase-1 enzyme which exists as a heterotetramer (homodimer of the P20 and P10 heterodimers) (2). Inflammasome complexes sense various microbial and endogenous signals, including ligand-receptor interaction and growth factor deprivation, which results in the processing of pro-caspase to active caspase-1 (3, 4).The active caspase-1 is responsible for the cleavage and maturation of pro-inflammatory cytokines IL-1beta and IL-18, and promotes an inflammatory form of cell death induced by bacterial pathogens called pyroptosis (3, 4). Mature IL-1beta has a role in the immune reaction and recruiting inflammatory cells to the site of infection while IL-18 plays a role in interferon-gamma production and promotes cytolytic activity in natural killer cells (4).

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 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Caspase-1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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