IL-27R alpha/WSX-1/TCCR Overexpression Lysate Summary
Description |
IL-27R alpha/WSX-1/TCCR Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC207012
Accession#: NM_004843
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. |
Gene |
IL27RA |
Applications/Dilutions
Dilutions |
|
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 |
69.3 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. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
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.
Alternate Names for IL-27R alpha/WSX-1/TCCR Overexpression Lysate
Background
Upon antigen challenge, T-helper cells differentiate into two functionally distinct subsets, Th1 and Th2. Th1 cells produce IL-2, IFN-g and lymphotoxin-b that augment cell mediated immune response while Th2 cells secrete IL-4, IL-5, and IL-10 that enhance humoral immunity. The function of T-helper cells is regulated by cytokines. A novel cytokine receptor was recently identified and cloned. It is a new member in the type I cytokine receptor family and designated TCCR for T-cell cytokine receptor and WSX-1. TCCR deficient mice had impaired Th1 responses to protein antigen challenge, including decreased levels of IFN-g and Th1-dependent antibody IgG2a. TCCR is predominantly expressed in thymus, spleen, lymph notes and peripheral blood leukocytes.
Limitations
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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