FAT10 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-17510
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for FAT10 Overexpression Lysate
FAT10 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC204431
Accession#: NM_006398
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.
Expression Host: HEK293T
Plasmid: RC204431
Accession#: NM_006398
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.
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
18.3 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.
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 FAT10 Overexpression Lysate
![Western Blot: FAT10 Overexpression Lysate [NBL1-17510]](https://resources.bio-techne.com/images/products/FAT10-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-17510-img0002.jpg "Western Blot: FAT10 Overexpression Lysate [NBL1-17510]")
Western Blot: FAT10 Overexpression Lysate [NBL1-17510]
Western Blot: FAT10 Overexpression Lysate (Adult Normal) [NBL1-17510] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for FAT10.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: FAT10
Human Leukocyte Antigen-F Associated Transcript 10 (FAT10), also known as Ubiquitin D (UBD), is a 165 amino acid (aa) member of the Ubiquitin-like family of proteins. Human FAT10 has a predicted molecular weight of 18.5 kDa and shares 69% aa sequence identity with mouse FAT10. Human FAT10 mRNA is expressed as a single transcript in lymphoblastoid lines and dendritic cells, but more than one mRNA transcript has been identified for murine FAT10. FAT10 can also be induced by IFN-gamma and TNF-alpha in some cell lines. Structurally, FAT10 consists of two Ubiquitin-like domains that are connected by a short linker. Like Ubiquitin, FAT10 has a C-terminal glycine residue that can be used to form isopeptide bonds with target proteins. FAT10-conjugated proteins are targeted to the proteasome where the 26S Proteasome subunit S5a/Angiocidin binds to FAT10 and enables subsequent degradation of the conjugated protein. In addition to S5a/Angiocidin, FAT10 has been shown to interact with Huntingtin, Ataxin-1, MAD2, and NUB1L. FAT10 has been implicated in a number of biological processes such as cell cycle control, antigen presentation, and cytokine response.
Alternate Names
UBD
Gene Symbol
UBD
Additional FAT10 Products
Product Documents for FAT10 Overexpression Lysate
Product Specific Notices for FAT10 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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