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CYLD Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-08356

Novus Biologicals, part of Bio-Techne
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NBP2-08356

Key Product Details

Species

Human

Applications

Functional Assay, Western Blot

Product Summary for CYLD Overexpression Lysate

CYLD Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC204099

Accession#: NM_001042355

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

106.9 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 CYLD Overexpression Lysate

Western Blot: CYLD Overexpression Lysate [NBP2-08356]

Western Blot: CYLD Overexpression Lysate [NBP2-08356]

Western Blot: CYLD Overexpression Lysate (Adult Normal) [NBP2-08356] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CYLD.

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: CYLD

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.

Long Name

Cylindromatosis

Alternate Names

BRSS, CDMT, CYLD1, EAC, MFT1, SBS, TEM, USPL2

Gene Symbol

CYLD

Additional CYLD Products

Product Documents for CYLD Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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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