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CNTF Overexpression Lysate

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Western Blot: CNTF Overexpression Lysate (Adult Normal) [NBP2-09006] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CNTF.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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CNTF Overexpression Lysate Summary

Description

CNTF Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC222331

Accession#: NM_000614

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.
Gene
CNTF

Applications/Dilutions

Dilutions
  • Western Blot
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
22.8 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.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

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.

Alternate Names for CNTF Overexpression Lysate

  • ciliary neurotrophic factor
  • CNTF
  • HCNTF

Background

CNTF is a survival promoting factor for different types of neurons in vitro and in vivo. The essential structural features for the biological function of human CNTF were investigated by Thier, M. et al. They showed that deletion of 14 N-terminal and 18 C-terminal amino acids significantly increased bioactivity compared to wild-type CNTF. FUNCTION: CNTF is a survival factor for various neuronal cell types. Seems to prevent the degeneration of motor axons after axotomy. SUBUNIT: Homodimer. SUBCELLULAR LOCATION: Cytoplasm. TISSUE SPECIFICITY: Nervous system. PHARMACEUTICAL: CNTF is being tested under the name Axokine by Regeneron Pharmaceuticals for treatment of human motor neuron diseases, such as amyotrophic lateral sclerosis (ALS). As it induces substantial weight loss, preferentially of fat as opposed to lean body mass, it is being used for obesity treatment. SIMILARITY: Belongs to the CNTF family.

Limitations

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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Product General Protocols

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol CNTF