Rat MYH3 cDNA Clone in Bacterial Expression Vector (His-MBP)
This is a Myosin Heavy Chain 3, Skeletal Muscle, Embryonic plasmid with cDNA insert cloned into Bacterial Expression VectorpPB-His-MBP. Insert length: 6035 bp. Transient expression. Suitable for Clon.
Bacterial selection: Kanamycin.
Catalog No. ABIN4749351
$6,532.84
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500 ng
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Quick Overview for Rat MYH3 cDNA Clone in Bacterial Expression Vector (His-MBP) (ABIN4749351)
Gene
MYH3
(Myosin Heavy Chain 3, Skeletal Muscle, Embryonic (MYH3))
Fusion tag
His-MBP
Application
Cloning (Clon)
Insert
cDNA
Vector
Bacterial Expression Vector
Vector Backbone
pPB-His-MBP
Promoter
T7 Promoter
Bacterial Resistance
Kanamycin
Expression Type
Transient
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Species
- Rat
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Purpose
- Bacterial expression of Rat Myh3 with His-MBP
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Specificity
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5-NheI and 3-XhoI
Fusion tag: Dual N-terminal tag, 6X Histidine followed by Maltose Binding Protein which is cleavable with Thrombin (Size 43 kDa) -
Insert Length
- 6035 bp
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Sequencing Primer
- MBP Forward primer: 5'-CGCAGATGTCCGCTTTCTGG-3', T7 terminator primer: 5'-GCTAGTTATTGCTCAGCGG-3'
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Application Notes
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The pPB vectors are low-medium copy number vectors in which the gene expression is driven by the strong T7 promoter.
Below are some basic guidelines for using the pPB vectors for protein production:
1. The pPB vectors are designed to be used with E. coli strains that are DE3 lysogens i.e. the host E. coli cell has a source of T7 RNA polymerase.
2. Recombinant protein induction is usually done at OD600 of 0.6-1.2 using Isopropyl β-D-1-thiogalactopyranoside (IPTG) at a final concentration of 0.05 -1mM.
3. The ideal concentration of IPTG must be determined empirically for each recombinant protein/cell-line. Similarly, the length of time and temperature for induction provide other variables that need to be optimized on a case-to-case basis.
4. For toxic proteins, it is recommended to go for shorter induction time and also to try and suppress basal recombinant gene expression through (a) addition of glucose or use of pLysS plasmid. Please note that special cell-lines are also available in the market that cater to expression of toxic proteins.
5. Once grown for the desired length of time, harvest cells by centrifugation and either freeze the cells at -80°C (as such or after re-suspending in the desired buffer) or proceed with the purification. -
Restrictions
- For Research Use only
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Format
- Liquid
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Buffer
- 10 mM Tris-HCI, 1 mM EDTA, pH 8.0
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Storage
- -20 °C
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Storage Comment
- 1 year when stored at -20° C or lower in a non-frost free freezer.
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Expiry Date
- 12 months
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- MYH3 (Myosin Heavy Chain 3, Skeletal Muscle, Embryonic (MYH3))
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Alternative Name
- Myh3
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NCBI Accession
- NM_012604
Target
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