Q: On the website, there is a WB image of this overexpressing lysate. Can you find out how much amount of the lysate was used for this WB?

A: The amount of lysate loaded in the western blot image was 5ug. We will generally recommend loading 5-10 ug per lane of the lysates in western blot analysis.

Q: What percentage of Fibulin5 is contained in this lysate?

A: Due to high volume of lysates been released, the blot on the website was probed with an anti-DDK antibody and not an anti-Fibulin 5. The exact concentration of the fibulin 5 protein in the lysates has not yet been determined. Concentrations are based on whole cell extract amounts.

Fibulin 5/DANCE Overexpression Lysate

Novus Biologicals | Catalog # NBL1-10609

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Fibulin 5/DANCE Overexpression Lysate

Fibulin 5/DANCE Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC204683

Accession#: NM_006329

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.

Theoretical MW

47.8 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.

Type

Overexpression

Scientific Data Images for Fibulin 5/DANCE Overexpression Lysate

Western Blot: Fibulin 5/DANCE Overexpression Lysate [NBL1-10609]

Western Blot: Fibulin 5 Overexpression Lysate (Adult Normal) [NBL1-10609] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Fibulin 5.

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: Fibulin 5/DANCE

The protein encoded by this gene is a secreted, extracellular matrix protein containing an Arg-Gly-Asp (RGD) motif and calcium-binding EGF-like domains. It promotes adhesion of endothelial cells through interaction of integrins and the RGD motif. It is prominently expressed in developing arteries but less so in adult vessels. However, its expression is reinduced in balloon-injured vessels and atherosclerotic lesions, notably in intimal vascular smooth muscle cells and endothelial cells. Therefore, the protein encoded by this gene may play a role in vascular development and remodeling. Defects in this gene are a cause of autosomal dominant cutis laxa, autosomal recessive cutis laxa type I (CL type I), and age-related macular degeneration type 3 (ARMD3). [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

Alternate Names

DANCE, EVEC, FBLN5, UP50

Gene Symbol

FBLN5

Additional Fibulin 5/DANCE Products

Product Documents for Fibulin 5/DANCE Overexpression Lysate

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Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Fibulin 5/DANCE 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.

Related Research Areas

Extracellular Matrix Molecules

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