ZIC1 Overexpression Lysate Summary
Description |
ZIC1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC211177
Accession#: NM_003412
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 |
ZIC1 |
Applications/Dilutions
Dilutions |
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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 |
48.1 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 ZIC1 Overexpression Lysate
Background
The Zic-1 gene, which encodes a zinc finger protein, is expressed in the developing or matured central nervous system in a highly restricted manner. The Zic gene is expressed in granule cells that make synaptic contact with Purkinje cells. Clearly Zic-1 is a gene critical to cerebellar pattern formation. The expression of Zic genes is first detected at gastrulation and at neurulation, becomes restricted to the dorsal neural ectoderm and the dorsal paraxial mesoderm. Zic-2 and Zic-3 are highly similar genes, especially in their product's zinc finger motif and by comparison of their genomic organization in that they share common exon-intron boundaries and belong to the same gene family. By comparison in function, Zic-2 is essential for the formation of the brain and Zic-3 is important for right and left axis formation. The Zic-1 gene has been mapped to chromosome 9 in mouse. The 5' flanking region of the Zic-1 gene contains a region-specific enhancer determined to be essential in in vivo and in vitro deletion analysis. The temporal profile of mRNA expression differs for each of the Zic gene products. The Drosophila odd-paired gene is highly homologous to the Zic gene family.
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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