Collagen I alpha 1 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-11172
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for Collagen I alpha 1 Overexpression Lysate
Collagen I alpha 1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC206234
Accession#: NM_000088
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: RC206234
Accession#: NM_000088
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
136.5 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 Collagen I alpha 1 Overexpression Lysate
Western Blot: Collagen I Overexpression Lysate (Adult Normal) [NBP2-11172]
Western Blot: Collagen I Overexpression Lysate (Adult Normal) [NBP2-11172] - Validation of overexpression lysate using anti-DDK antibody. Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC206234 using transfection reagent.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: Collagen I alpha 1
Type I collagen is a fibril-forming collagen found in most connective tissues and is abundant in bone, cornea, dermis and tendon tissue. Collagens are fibrous, extracellular matrix proteins with high tensile strength and are the major components of connective tissue. Several collagens play a role in cell adhesion, responsible for maintaining normal tissue architecture and function. All collagens contain a triple helix domain and frequently show lateral self-association in order to form complex connective tissues. Post-Golgi LH3 trafficking is essential for collagen homeostasis and for the development and function of multiple organs and tissues (1).
The COL1A1 gene encodes the pro-alpha1 chains of type I collagen protein, whose triple helix is comprised of two alpha1 chains and one alpha2 chain. Mutations in the encoding COL1A1 gene are associated with brittle bone disease (Osteogenesis Imperfecta), cortical hyperostosis (Caffey disease) and disorders that affect the connective tissues (Ehlers-Danlos syndrome) (2). Studies have found that HIF-1 transcription regulation of collagen prolyl hydroxylases regulates collagen deposition, promoting cancer cell alignment along collagen fibers, which enhances invasion and metastasis to lymph nodes and lung tissue by breast cancer cells (3).
References
1. Banushi, B., Forneris, F., Straatman-Iwanowska, A., Strange, A., Lyne, A. M., Rogerson, C., . . . Gissen, P. (2016). Regulation of post-Golgi LH3 trafficking is essential for collagen homeostasis. Nat Commun, 7, 12111. doi:10.1038/ncomms12111
2. Lu, Y., Zhang, S., Wang, Y., Ren, X., & Han, J. (2019). Molecular mechanisms and clinical manifestations of rare genetic disorders associated with type I collagen. Intractable Rare Dis Res, 8(2), 98-107. doi:10.5582/irdr.2019.01064
3. Gilkes, D. M., Chaturvedi, P., Bajpai, S., Wong, C. C., Wei, H., Pitcairn, S., . . . Semenza, G. L. (2013). Collagen prolyl hydroxylases are essential for breast cancer metastasis. Cancer Res, 73(11), 3285-3296. doi:10.1158/0008-5472.Can-12-3963
Alternate Names
COL1A1, OI4
Gene Symbol
COL1A1
Additional Collagen I alpha 1 Products
Product Documents for Collagen I alpha 1 Overexpression Lysate
Product Specific Notices for Collagen I alpha 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.
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