Ly-6G6C Overexpression Lysate Summary
Description |
Ly-6G6C Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC216027
Accession#: NM_025261
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 |
LY6G6C |
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 |
13.6 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 Ly-6G6C Overexpression Lysate
Background
Ly6G is a GPI-anchored protein, that is also known as the myeloid differentiation antigen Gr1. The antigen is transiently expressed on monocytes in the bone marrow. The level of antigen expression in the bone marrow directly correlates with granulocyte differentiation and maturation. Ly6G is expressed predominantly on neutrophils, also in a subset of eosinophils, differentiating pre-monocytes, and plasmacytoid dendritic cells.
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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Product General Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
Video Protocols
FAQs for Ly-6G6C Lysate (NBP2-06220). (Showing 1 - 1 of 1 FAQ).
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I am running this lysate on a western as a positive control. Approximately what amount/volume is run on the gel?
- Generally for our over-expression lysates, 10-25ug is recommended for loading on a gel as a positive control for the first time. For this particular lysate 10ug was tested first, and since a clear signal was seen, this image was used on the website.
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