alpha-2A Adrenergic R/ADRA2A Overexpression Lysate

Western Blot: alpha-2A Adrenergic R/ADRA2A Overexpression Lysate [NBL1-07357] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Alpha 2a Adrenergic Receptor.

• Reactivity: Hu
• Applications: WB

alpha-2A Adrenergic R/ADRA2A Overexpression Lysate Summary

• Description:

  alpha-2A Adrenergic R/ADRA2A Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC208366
  
  Accession#: NM_000681
  
  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: ADRA2A

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: 48.8 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 alpha-2A Adrenergic R/ADRA2A Overexpression Lysate

• a-2A AdrenergicR
• ADRA2
• ADRA2A
• ADRA2Ralpha-2-adrenergic receptor, platelet type
• ADRARZNF32
• adrenergic, alpha-2A-, receptor
• Alpha-2 adrenergic receptor subtype C10
• alpha-2A Adrenergic R
• alpha-2A adrenergic receptor
• alpha2A AdrenergicR
• Alpha-2A adrenoceptor
• Alpha-2A adrenoreceptor
• alpha-2AAR subtype C10
• ALPHA2AAR
• Alpha-2AAR
• ZNF32

Background

Alpha-2-adrenergic receptors are members of the G protein-coupled receptor superfamily. They include 3 highly homologous subtypes: alpha2A, alpha2B, and alpha2C. These receptors have a critical role in regulating neurotransmitter release from sympathetic nerves and from adrenergic neurons in the central nervous system. Studies in mouse revealed that both the alpha2A and alpha2C subtypes were required for normal presynaptic control of transmitter release from sympathetic nerves in the heart and from central noradrenergic neurons; the alpha2A subtype inhibited transmitter release at high stimulation frequencies, whereas the alpha2C subtype modulated neurotransmission at lower levels of nerve activity. This gene encodes alpha2A subtype and it contains no introns in either its coding or untranslated sequences. [provided by RefSeq]. Sequence Note: The 5'-most in-frame translation initiation codon is selected for this RefSeq based on good conservation across mammalian species. A possible downstream start codon would result in a protein that is 15 aa shorter at the N-terminus. The literature, including PMIDs:2823383, 2568356, 1354394 and 1678390, assumes the use of the downstream start codon based on initial cloning reports, but there is no experimental evidence indicating which start codon is preferentially used in vivo. CCDS Note: The coding region has been updated to extend the N-terminus to one that is more supported by available transcript and conservation data. Two possible start codons are present in this update, with each having a weak Kozak signal. Due to leaky scanning by ribosomes, it is possible that some ribosomes may initiate translation from the upstream AUG while others start from the downstream AUG. Translation initiation from the downstream AUG would result in a protein that is 15 aa shorter at the N-terminus. The literature, including PMIDs 2823383, 2568356, 1354394 and 1678390, assumes the use of the downstream start codon based on initial cloning reports, but there is no experimental evidence indicating which start codon is preferentially used in vivo. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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: ADRA2A