mGluR4 Overexpression Lysate

Western Blot: metabotropic Glutamate Receptor 4 Overexpression Lysate (Adult Normal) [NBL1-11349] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for metabotropic Glutamate Receptor 4.

• Reactivity: Hu
• Applications: WB

mGluR4 Overexpression Lysate Summary

• Description:

  mGluR4 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC223673
  
  Accession#: NM_000841
  
  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: GRM4

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: 98.2 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 mGluR4 Overexpression Lysate

• glutamate receptor, metabotropic 4
• GPRC1D
• GPRC1Dmetabotropic glutamate receptor 4
• GRM4
• MGC177594
• mGlu4
• mGluR4
• MGLUR4glutamate metabotropic receptor 4

Background

The Metabotropic Glutamate Receptor mGluR4/GRM4 binds L-glutamate, the major excitatory neurotransmitter in the central nervous system, which activates both ionotropic and metabotropic glutamate receptors. Glutamatergic neurotransmission is involved in most aspects of normal brain function and can be perturbed in many neuropathologic conditions. The metabotropic glutamate receptors are a family of G protein-coupled receptors that have been divided into 3 groups on the basis of sequence homology, putative signal transduction mechanisms, and pharmacologic properties. Group III includes GRM4, GRM6, GRM7, and GRM8 receptors that are linked to the inhibition of the cyclic AMP signaling cascade. Two protein isoforms have been reported in rat (mGluR4a 912 aa; mGluRb 983 aa), which differ at the C-terminus. GRM4 has been reported in various regions of the brain, eye, and spinal cord. ESTs have been isolated from brain, embryo, melanocyte/uterus/fetal heart, nerve, and testis libraries. Caution: GLUR4 refers to Ionotropic Glutamate Receptor 4, not to Metabotropic Glutamate Receptor 4 (mGLUR4).

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: GRM4