PMCA4 Overexpression Lysate Summary
Description |
PMCA4 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC213835
Accession#: NM_001684
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 |
ATP2B4 |
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 |
133.8 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 PMCA4 Overexpression Lysate
Background
The protein encoded by the PMCA4 gene belongs to the family of P-type primary ion transport ATPases characterized by theformation of an aspartyl phosphate intermediate during the reaction cycle. These enzymes remove bivalent calcium ionsfrom eukaryotic cells against very large concentration gradients and play a critical role in intracellular calciumhomeostasis. The mammalian plasma membrane calcium ATPase isoforms are encoded by at least four separate genes and thediversity of these enzymes is further increased by alternative splicing of transcripts. The expression of differentisoforms and splice variants is regulated in a developmental, tissue- and cell type-specific manner, suggesting thatthese pumps are functionally adapted to the physiological needs of particular cells and tissues. This gene encodes theplasma membrane calcium ATPase isoform 4. Alternatively spliced transcript variants encoding different isoforms havebeen identified. (provided by RefSeq)
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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