Ubiquitin carboxyl-terminal hydrolase CYLD (CYLD) is a 956 amino acid (aa) member of the peptidase C67 protein family with a predicted molecular weight of 107 kDa. The mouse and rat CYLD orthologs share 95% and 94% aa sequence identity with the human protein, respectively. Two isoforms of CYLD have been identified, a full-length isoform and a second isoform that lacks aa 305-307 due to alternative splicing. Expression of CYLD has been reported in fetal brain as well as adult brain, heart, leukocytes, skeletal muscle, spleen, and testis. CYLD acts as a deubiquitinase and removes K63-linked Ubiquitin chains from multiple substrates including IkB, c-Jun, and c-Fos, resulting in the inhibition of NFkB and JNK signaling. In some contexts, CYLD enhances mitosis entry, and it has also been shown to delay G 1 /S phase entry suggesting that CYLD regulates multiple phases of the cell cycle. CYLD is recognized as a tumor suppressor and mutations in CYLD result in skin appendage syndromes including Brooke-Spiegle Syndrome, familial cylindromatosis, and familial trichoepitheliomas type 1.
CYLD Overexpression Lysate
Novus Biologicals | Catalog # NBP2-08360
Key Product Details
Species
Applications
Product Summary for CYLD Overexpression Lysate
CYLD Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC214451
Accession#: NM_001042412
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.
Theoretical MW
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
Scientific Data Images for CYLD Overexpression Lysate
Western Blot: CYLD Overexpression Lysate [NBP2-08360]
Western Blot: CYLD Overexpression Lysate (Adult Normal) [NBP2-08360] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CYLD.Formulation, Preparation, and Storage
Formulation
Concentration
Shipping
Storage
Background: CYLD
Long Name
Alternate Names
Gene Symbol
Additional CYLD Products
Product Documents for CYLD Overexpression Lysate
Certificate of Analysis
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Product Specific Notices for CYLD 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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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars