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Connexin 32/GJB1 Overexpression Lysate

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Western Blot: GJB1 Overexpression Lysate (Adult Normal) [NBP2-08276] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GJB1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Connexin 32/GJB1 Overexpression Lysate Summary

Description

Connexin 32/GJB1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC211786

Accession#: NM_001097642

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
GJB1

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 (cat# 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
31.8 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 Connexin 32/GJB1 Overexpression Lysate

  • CMTX
  • CMTX1
  • Connexin 32
  • Connexin-32
  • CX32
  • CX32beta 1, 32kD (connexin 32, Charcot-Marie-Toothneuropathy, X-linked)
  • gap junction beta-1 protein
  • gap junction protein, beta 1, 32kDa (connexin 32)
  • gap junction protein, beta 1, 32kDa (connexin 32, Charcot-Marie-Toothneuropathy, X-linked)
  • gap junction protein, beta 1, 32kDa
  • GJB1

Background

Gap junctions are aggregations of intercellular channels that directly connect the cytoplasm of adjacent cells. Gap junctions coordinate cellular and organ function in tissues and are involved in metabolic cooperation between cells, synchronization of cellular physiological activities, growth control and developmental regulation. The gap junction channels allow intercellular exchange of ions, nucleotides and small molecules between adjacent cells. Unlike other membrane channels, intercellular channels span two plasma membranes and require the contribution of hemichannels, called connexons, from both participating cells. These channels are permeate to molecules as large as 1 kD, and they have been detected in virtually every cell type in mammals, except mature skeletal muscle, spermatozoa and erythrocytes. Two connexons interact in the extracellular space to form the complete intercellular channel. Each connexon is composed of six similar or identical proteins, which are termed connexins. Connexins (Cx) are a multi-gene family of highly related proteins ranging between molecular weights of 26 and 70 kD. At least a dozen distinct connexin genes have been identified and many are expressed in a tissue-specific manner. Two distinct lineages have been identified in mammals. One termed class I or beta group to which Cx26, Cx30, Cx31, Cx31.1 and Cx32 belong, and the other termed class II or alpha group, represented by Cx33, Cx37, Cx40, Cx43 and Cx46.

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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Bioinformatics

Gene Symbol GJB1