TDAG8/GPR65 Overexpression Lysate

Western Blot: GPR65 Overexpression Lysate (Adult Normal) [NBP2-04251] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GPR65.

• Reactivity: Hu
• Applications: WB

TDAG8/GPR65 Overexpression Lysate Summary

• Description:

  TDAG8/GPR65 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC207600
  
  Accession#: NM_003608
  
  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: GPR65

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: 39.2 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.

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 TDAG8/GPR65 Overexpression Lysate

• G protein-coupled receptor 65
• GPR65
• G-protein coupled receptor 65
• Psychosine Receptor
• T-cell death-associated gene 8 protein
• TDAG8
• TDAG8hTDAG8

Background

GPR65 (TDAG8), a Lysophospholipid/Lysosphingolipid Receptor, is a molecular target for psychosine, a toxic lipid formed by the breakdown of galactosylceramide (cerebroside). A build-up of psychosine, caused by the lack of galactosylceramidase, results in the death of the myelin-synthesizing cells, oligodendrocytes, which is associated with Krabbe disease. GPR65 has been reported in human in peripheral blood leukocytes, spleen, lymph node, and thymus. ESTs have been isolated from human B-cell/lung/testis, brain/lung/testis, pancreas/spleen, lung/spleen, leukocyte, embryo, kidney, liver/spleen, thyroid, and vessel libraries.

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.

FAQs for TDAG8/GPR65 Lysate (NBP2-04251). (Showing 1 - 1 of 1 FAQ).

1. Has the anti-GPR65 antibody, NBP2-24487, been tested in the overexpressing lysate, NBP2-04251?
   • We do not have data from the tests if indeed these have been used together, but it would make sense to utilize the lysate as a positive control for your western blot if needed.

Bioinformatics

• Gene Symbol: GPR65