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Human Sonic Hedgehog/SHH HEK293 Overexpression Lysate

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Specification

Cat. No.

NSC1123

Category

Neural Stem Cell Growth Factors

Product Type

Lysates

Product Name

Human Sonic Hedgehog/SHH HEK293 Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human HHG1 Overexpression Lysate; Human HLP3 Overexpression Lysate; Human HPE3 Overexpression Lysate; Human MCOPCB5 Overexpression Lysate; Human SMMCI Overexpression Lysate; Human Sonic hedgehog Overexpression Lysate; Human TPT Overexpression Lysate; Human TPTPS Overexpression Lysate

Species

Human

Molecule Mass

The recombinant human SHH( aa 198-462) consists of 276 amino acids and has a predicted molecular mass of 29 kDa. The apparent molecular mass of the protein is approximately 35 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Stability & Storage

Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.

Product Description

This Human Sonic Hedgehog/SHH overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Sonic Hedgehog/SHH protein (Cat: 10372-H08H) from the overexpression lysate was verified.

Expression Host

HEK293 Cells

Sequence Information

A DNA sequence encoding the amino acid sequence (Cys 198-Ser 462) of human SHH (Q15465), that is Sonic hedgehog protein C-product, was fused with a polyhistidine tag at the C-terminus and a signal peptide at the N-terminus.

Preparation Method

Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.

Lysis Buffer

Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.

Recommend Usage

1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.

Sample Buffer

1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

Application

Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.