Sonic Hedgehog/Shh Overexpression Lysate

Name: Sonic Hedgehog/Shh Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-15941

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-15941

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

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Sonic Hedgehog/Shh Overexpression Lysate

Sonic Hedgehog/Shh Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC222175

Accession#: NM_000193

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.

TMW

47.2 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 Sonic Hedgehog/Shh Overexpression Lysate

Western Blot: Sonic Hedgehog/Shh Overexpression Lysate [NBL1-15941]

Western Blot: Sonic Hedgehog Overexpression Lysate (Adult Normal) [NBL1-15941] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Sonic Hedgehog.

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: Sonic Hedgehog/Shh

Sonic Hedgehog Protein (SHH, VHH-1) belongs to hedgehog protein family which includes Indian Hh, and Desert Hh. Hh family is involved in the cell fate and patterning during embryonic development, homeostasis, and adult tissue renewal (1). Similar to other member in the family, SHH binds to the patched (PTC) cell surface receptor, releasing the signal transducer Smoothened (Smo) to transmit the Hh signal into the cell and activate transcription of the target gene (2). Precursor SHH is autocatlytically cleaved into two subunits, N-terminal and C-terminal products. Soluble N-terminal product is involved in the signaling activity, while C-product displays an autoproteolysis activity on the precursor and a cholesterol transferase activity on the N-terminal product. C-terminal product attaches a cholesterol moiety to the N-terminal product, preventing N-terminal product diffusion within the developing embryo (3). A defect in SHH has been linked in holoprosencephaly type 3 (HPE3), in which developing forebrain fails to separate into right and left hemispheres, and ocular coloboma (4).

Alternate Names

HHG1, HLP3, HPE3, MCOPCB5, Shh, ShhNC, SMMCI, TPTPS

Gene Symbol

SHH

Additional Sonic Hedgehog/Shh Products

Product Documents for Sonic Hedgehog/Shh Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Sonic Hedgehog/Shh 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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