SIRP alpha/CD172a Overexpression Lysate Summary
Description |
SIRP alpha/CD172a Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC216320
Accession#: NM_001040023
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 |
SIRPA |
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 (cat# 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 |
52.3 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 SIRP alpha/CD172a Overexpression Lysate
Background
Protein tyrosine phosphatases (PTPases) SHP-1 and SHP-2 are critical regulators in the intracellular signaling pathways that result in cell responses such as mitosis, differentiation, migration, survival, transformation or death. SHP-2 is a signal transducer for several receptor tyrosine kinases and cytokine receptors. A novel SHP-2 associated glycoprotein was recently cloned from human, rat, mouse and cattle by several labs and was designated SIRPa (1), SHPS-1 (2,3), MyD-1 (4), BIT (5,6) and p84 (7). SIRPa is a new gene family containing at least fifteen members. SIRPa is a substrate of many activated tyrosine kinases such as insulin receptor, EGFR, PDGFR and src, and a specific docking protein for SHP-2 (1,2,5,8). SIRPa has regulatory effects on cellular responses induced by serum, growth factors, insulin, oncogenes, growth hormones and cell adhesion and plays a general role in different physiological and pathological processes (1,2,5,8).
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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