SP2 Overexpression Lysate Summary
| Description |
SP2 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC207602
Accession#: NM_003110
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 |
SP2 |
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 |
64.7 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 SP2 Overexpression Lysate
Background
The Sp transcription factor family includes Sp1, Sp2, Sp3 (SPR-2) and Sp4 (SPR-1). Sp transcription factors share similar structures but do not share similar functions. All four proteins contain a highly conserved DNA-binding domain composed of three zinc fingers at the C-terminus. Sp family members bind the consensus sequence GGGGCGGGGC and other closely related sequences which are known as GC boxes. Sp1, Sp3 and Sp4 share a high affinity for GC boxes while Sp2 does not. Sp2 only weakly binds to GT boxes. Sp1, Sp2 and Sp3 are ubiquitously expressed, while Sp4 is abundantly expressed in brain with limited expression in other tissues. Sp1 and Sp3, but not Sp2 or Sp4, interact with E2, a regulatory element for the b4 subunit of neuronal nicotinic acetylcholine receptors. Sp3 is the only Sp member to inhibit Sp1 and Sp4 mediated transcription. The gene3 encoding human Sp2 maps to chromosome 17p32.3.
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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