Connexin 50/GJA8 Overexpression Lysate

Western Blot: GJA8 Overexpression Lysate (Adult Normal) [NBP2-07766] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GJA8.

• Reactivity: Hu
• Applications: WB

Connexin 50/GJA8 Overexpression Lysate Summary

• Description:

  Connexin 50/GJA8 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC213469
  
  Accession#: NM_005267
  
  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: GJA8

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: 48 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 50/GJA8 Overexpression Lysate

• alpha 8, 50kD (connexin 50)
• CAE
• CAE1
• Connexin 50
• Connexin-50
• CX50
• CZP1
• gap junction alpha-8 protein
• gap junction membrane channel protein alpha-8
• gap junction protein, alpha 8, 50kDa (connexin 50)
• gap junction protein, alpha 8, 50kDa
• GJA8
• lens intrinsic membrane protein MP70
• MP70

Background

The 433 amino acid long, 48 kDA transmembrane connexin, gap junction alpha-8 protein encoded by the GJA8 gene is critical in growth and development of lens fiber cells. Mutations in the GJA8 gene may lead to zonular pulverulent cataracts, nuclear progressive cataracts, and cataract-microcornea syndrome. The GJA8 gene has been linked to myopia, microphthalmia, neuronitis, cycstic fibrosis, neuopathy, cervical adenocarcinoma, and juvenile myoclonic epilepsy. The GJA8 gene interacts with TJP1, MIP, MED14, GJA1, and GJB1 to participate in pathways such as gap junction assembly, membrane trafficking, and connexin synthesis.

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.

Bioinformatics

• Gene Symbol: GJA8