CARD12 Overexpression Lysate

Name: CARD12 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-06545

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06545

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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 CARD12 Overexpression Lysate

CARD12 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC206757

Accession#: NM_021209

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

116 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 CARD12 Overexpression Lysate

Western Blot: CARD12 Overexpression Lysate [NBP2-06545]

Western Blot: CARD12 Overexpression Lysate (Adult Normal) [NBP2-06545] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CARD12.

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: CARD12

Ipaf (also known as Clan/CARD12) is a CARD domain containing protein. CARD (caspase-associated recruitment domain) proteins are key regulators of cell death, cell survival and cytokine production (reviewed in Damiano and Reed, 2004). In general CARD proteins are implicated in host defense against infection, environmental stress or cellular damage. CARD domains are found in the N-terminal pro-domains of certain caspases, a family of apoptotic and pro-inflammatory proteases, as well as in a diversity of other proteins including Ipaf/Clan/CARD12. CARD domains are homotypic protein interaction motifs that enable networks of proteins to communicate via CARD-CARD interactions. There are at least three major signaling pathways in which CARD proteins act: (1) Regulation of caspase activation in the context of apoptosis (2) Regulation of caspase activation in the context of inflammation (3) Regaultion of NF-kB activation in the context of innate or adaptive immune responses. As there is significant crosstalk between pathways that lead to caspase-mediated apoptosis or inflammation and pathways that result in NF-kB activation, it is logical that similar protein modules such as CARD domains are found repeatedly in proteins from all three pathways. Ipaf plays a role in regulating caspase-1 activity, which in turn mediates the maturation of inflammatory cytokines IL-1b and IL-18 (reviewed in Lu et al, 2005). In transfected cells, Ipaf has been shown to directly interact with procaspase-1 and induce proteolytic activation of procaspase-1 in transfected cells. On the flip side, macrophages from IPAF deficient mice failed to activate caspase-1 in response to Salmonella typhimurium infection underscoring the importance of IPAF in vivo. IPAF also interact with the pro-apoptotic adaptor protein ASC and co-expression of IPAF with ASC has been shown to induce NF-kB activation and apoptosis.

Alternate Names

CARD12leucine rich repeat and CARD domaincontaining 4, caspase recruitment domain family, member 12, Caspase recruitment domain-containing protein 12, Clan protein, CLAN1ipaf, CLANCARD, LRR, and NACHT-containing protein, CLR2.1, Ice protease-activating factor, Ipaf, NLR family, CARD domain containing 4

Gene Symbol

NLRC4

Additional CARD12 Products

Product Documents for CARD12 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for CARD12 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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