PGLYRP1/PGRP-S Overexpression Lysate Summary
| Description |
PGLYRP1/PGRP-S Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC211040
Accession#: NM_005091
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 |
PGLYRP1 |
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 |
21.6 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 PGLYRP1/PGRP-S Overexpression Lysate
Background
The primary immune recognition is based on structures common among invading pathogens. Bacterial surface molecules, such as lipopolysaccharide (LPS) and peptidoglycan (PGN), are known to elicit immune reactions ranging from cytokine release to fever. Recently, a family of proteins called peptidoglycan recognition protein (PGRP) has been identified in mouse and human that binds to peptidoglycans expressed on Gram-positive bacteria. Peptidoglycan (PGN) is an essential cell wall component of virtually all bacteria (1,2) and, thus, it is an excellent target for recognition by the eukaryotic innate immune system. The PGRPs (PGRP-L, PGRP-S, PGRP-Ia, and PGRP-Ib) define a new family of human pattern recognition molecules (3). PGRP-L is primarily expressed in the liver. Although liver is not considered a primary immune organ, liver participates in host defenses by producing acute phase proteins (by hepatocytes) in response to infections and by clearing microorganisms from blood (4). PGRP-S is present in neutrophils and inhibits growth of Gram-positive bacteria and, therefore, may function as a neutrophil antibacterial protein (5). However, PGRP-S may have another, as yet unidentified function because in humans it is expressed in the bone marrow 50
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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