ASC/TMS1 Overexpression Lysate

Name: ASC/TMS1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-15016

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-15016

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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 ASC/TMS1 Overexpression Lysate

ASC/TMS1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC215592

Accession#: NM_013258

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

21.4 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 ASC/TMS1 Overexpression Lysate

Western Blot: ASC/TMS1 Overexpression Lysate [NBL1-15016]

Western Blot: ASC/TMS1 Overexpression Lysate (Adult Normal) [NBL1-15016] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ASC/TMS1.

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: ASC

ASC (apoptosis-associated speck-like protein containing a CARD), also known as TMS1 (target of methylation-induced silencing), was first identified in 1999 as a protein that forms aggregates, or specks, during retinoic acid-induced apoptosis in a human leukemia cell line (1). Furthermore, it was discovered to have a role as a tumor suppressor as methylation silences ASC/TMS1 expression in many tumors (2-5). ASC/TMS1 is synthesized as a 195-amino acid (aa) protein with a theoretical weight of 22 kDa. Structurally the protein contains a N-terminal PYD (pyrin domain) and C-terminal CARD (caspase-recruitment domain) (1-4). Historically, CARD and PYD-containing proteins are known to have crucial functions in regulating apoptosis and immune response pathways (2-5). Furthermore, mutations in many CARD and PYD-containing proteins have been linked to various cancers and inflammatory diseases (2-5). Given its immune response role, it is not surprising that ASC/TMS1 is typically highly expressed in immune cells, specifically in neutrophils and cells of the macrophage/monocyte lineage (5). Additionally, it is expressed in many normal epithelial cell types (4,5).

In regard to immune and inflammatory response, ASC/TMS1 is involved in inflammasome function (3-4). The inflammasome is a multiprotein complex that responds to cellular stress or pathogens and activates inflammatory responses. Specifically, ASC/TMS1 helps assemble the NLRP3 inflammasome complex which then activates caspase-1, followed by stimulation of proinflammatory cytokines including IL-1b and IL-18 (3-4). In terms of the role in regulating apoptosis, multiple studies have revealed that the ASC/TMS1 gene is hypermethylated in many cancers including breast, lung, glioblastomas, and melanomas (2-5). The increased methylation results in decreased gene expression, or silencing, allowing those cancer cells to escape apoptosis (2-5).

References

1. Masumoto, J., Taniguchi, S., Ayukawa, K., Sarvotham, H., Kishino, T., Niikawa, N., Hidaka, E., Katsuyama, T., Higuchi, T., & Sagara, J. (1999). ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. The Journal of biological chemistry, 274(48), 33835-33838. https://doi.org/10.1074/jbc.274.48.33835

2. McConnell, B. B., & Vertino, P. M. (2004). TMS1/ASC: the cancer connection. Apoptosis: an international journal on programmed cell death. https://doi.org/10.1023/B:APPT.0000012117.32430.0c

3. Salminen, A., Kauppinen, A., Hiltunen, M., & Kaarniranta, K. (2014). Epigenetic regulation of ASC/TMS1 expression: potential role in apoptosis and inflammasome function. Cellular and molecular life sciences : CMLS. https://doi.org/10.1007/s00018-013-1524-9

4. Protti, M. P., & De Monte, L. (2020). Dual Role of Inflammasome Adaptor ASC in Cancer. Frontiers in cell and developmental biology. https://doi.org/10.3389/fcell.2020.00040

5. Parsons, M. J., & Vertino, P. M. (2006). Dual role of TMS1/ASC in death receptor signaling. Oncogene. https://doi.org/10.1038/sj.onc.1209684

Long Name

Apoptosis-Associated Speck-Like Protein Containing A CARD

Alternate Names

CARD5, PYCARD, TMS1

Gene Symbol

PYCARD

Additional ASC Products

Product Documents for ASC/TMS1 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for ASC/TMS1 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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