Androgen R/NR3C4 Overexpression Lysate Summary
Description |
Androgen R/NR3C4 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC215316
Accession#: NM_000044
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 |
AR |
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 |
99 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 Androgen R/NR3C4 Overexpression Lysate
Background
Androgens have a broad range of effects on the appearance and maintenance of male secondary sexual characteristics. Like other members of the steroid superfamily, the androgen receptor (AR) consists of an amino terminal modulating domain, a central DNA binding domain, a hinge region and a carboxy terminal ligand binding domain. The androgen receptor (AR) is a 110 kDa androgen-dependent transcription factor that is a member of the steroid/nuclear receptor gene superfamily. The AR signaling pathway plays a key role in development and function of male reproductive organs, including the prostate and epididymis. AR also plays a role in nonreproductive organs, such as muscle, hair follicles, and brain. Abnormalities in the AR signaling pathway have been linked to a number of diseases, including prostate cancer, Kennedy's disease and male infertility. The PI3K/Akt signaling pathway plays an important role in regulating AR activity through phosphorylation of AR at Ser213/210 and Ser791/790. Growth factors or cytokines may induce phosphorylation of AR through the PI3K/Akt pathway. Activation of the PI3K/AKt pathway is thought to have a survival role in prostate cancer by protecting cells from apoptosis.
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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