CD31/PECAM-1 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10670
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for CD31/PECAM-1 Overexpression Lysate
CD31/PECAM-1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC208654
Accession#: NM_000442
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.
Expression Host: HEK293T
Plasmid: RC208654
Accession#: NM_000442
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.
Product Specifications
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.
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.
TMW
82.5 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.
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.
Type
Overexpression
Scientific Data Images for CD31/PECAM-1 Overexpression Lysate
![Western Blot: CD31/PECAM-1 Overexpression Lysate [NBP2-10670]](https://resources.bio-techne.com/images/products/CD31-PECAM-1-Overexpression-Lysate-Western-Blot-NBP2-10670-img0001.jpg "Western Blot: CD31/PECAM-1 Overexpression Lysate [NBP2-10670]")
Western Blot: CD31/PECAM-1 Overexpression Lysate [NBP2-10670]
Western Blot: CD31/PECAM-1 Overexpression Lysate [NBP2-10670] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -CD31/PECAM-1 Overexpression Lysate.Formulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: CD31/PECAM-1
PECAM's intracellular cytoplasmic domain consists of a sequence of 118 amino acids and contains serine and tyrosine (also referred to as immunoreceptor tyrosine-based inhibitory motifs-ITIMs) residues, which may be phosphorylated upon cellular stimulation (3). ITIMs are phosphorylated by Src-family kinases and non-Src family kinases (e.g., Csk), leading to a conformational change which supports interactions with Src homology 2 (SH2) domain containing proteins such as protein-tyrosine phosphatase, SHP-2 (1,2). Formation of SHP-2/PECAM-1 complexes induces endothelial cell migration through the dephosphorylation of focal adhesion kinase and regulation of RhoA activity (1). Signaling downstream of ITIM tyrosine phosphorylations also plays a role in PECAM's anti-apoptotic activity, a function which is independent of its interaction with SHP-2. In platelets and leukocytes, phosphorylation of PECAM's cytosolic domain is inhibitory, preventing their activation.
References
1. Lertkiatmongkol, P., Liao, D., Mei, H., Hu, Y., & Newman, P. J. (2016). Endothelial functions of PECAM-1 (CD31). Current Opinion in Hematology. https://doi.org/10.1097/MOH.0000000000000239.Endothelial
2. Privratsky, J. R., & Newman, P. J. (2014). PECAM-1: Regulator of endothelial junctional integrity. Cell and Tissue Research. https://doi.org/10.1007/s00441-013-1779-3
3. Newman, P. J., & Newman, D. K. (2003). Signal transduction pathways mediated by PECAM-1: New roles for an old molecule in platelet and vascular cell biology. Arteriosclerosis, Thrombosis, and Vascular Biology. https://doi.org/10.1161/01.ATV.0000071347.69358.D9
Long Name
Platelet Endothelial Cell Adhesion Molecule 1
Alternate Names
CD31, EndoCAM, PECA1, PECAM-1, PECAM1
Gene Symbol
PECAM1
Additional CD31/PECAM-1 Products
Product Documents for CD31/PECAM-1 Overexpression Lysate
Product Specific Notices for CD31/PECAM-1 Overexpression Lysate
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.
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.
Loading...
Loading...
Loading...
Loading...