Q: How much Butyrylcholinesterase lysate[NBL1-07942] do you use in the Butyrylcholinesterase Lysate Images? (What amount of protein should I load for overexperssion lysates?)

A: The lysate WB images you inquired about show 10ug of transfected lysate and 10ug of empty vector control lysate being loaded per well. (We recommend loading 10ug of protein per lane of our overexpression lysates. The end user will need to determine their own optimal working amounts.)

Butyrylcholinesterase/BCHE Overexpression Lysate

Novus Biologicals | Catalog # NBL1-07942

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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 Butyrylcholinesterase/BCHE Overexpression Lysate

Butyrylcholinesterase/BCHE Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC202198

Accession#: NM_000055

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

68.2 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 Butyrylcholinesterase/BCHE Overexpression Lysate

Western Blot: Butyrylcholinesterase/BCHE Overexpression Lysate [NBL1-07942]

Western Blot: Butyrylcholinesterase/BCHE Overexpression Lysate [NBL1-07942] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Butyrylcholinesterase.

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: Butyrylcholinesterase/BCHE

Mutant alleles at the BCHE locus are responsible for suxamethonium sensitivity. Homozygous persons sustain prolonged apnea after administration of the muscle relaxant suxamethonium in connection with surgical anesthesia. The activity of pseudocholinesterase in the serum is low and its substrate behavior is atypical. In the absence of the relaxant, the homozygote is at no known disadvantage. [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

Acylcholine acylhydrolase, BCHE, Butyrylcholine esterase, CHE1, Pseudocholinesterase

Gene Symbol

BCHE

Additional Butyrylcholinesterase/BCHE Products

Product Documents for Butyrylcholinesterase/BCHE 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.

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Product Specific Notices for Butyrylcholinesterase/BCHE 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

Neurotransmitters, Neuroactive Molecules and Associated Enzymes

Phosphatases and Regulators

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