HCN2 Overexpression Lysate

Western Blot: HCN2 Overexpression Lysate (Adult Normal) [NBP2-08224] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HCN2.

• Reactivity: Hu
• Applications: WB

HCN2 Overexpression Lysate Summary

• Description:

  HCN2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC214036
  
  Accession#: NM_001194
  
  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: HCN2

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: 96.8 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 HCN2 Overexpression Lysate

• BCNG2
• BCNG-2
• brain cyclic nucleotide gated channel 2
• brain cyclic nucleotide-gated channel 2
• HAC-1
• HCN2
• hyperpolarization activated cyclic nucleotide-gated potassium channel 2
• potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 2

Background

Hyperpolarization-activated cation channels of the HCN gene family such as HCN2, contribute to spontaneous rhythmic activity in both heart and brain. HCN2 is a member of a family of pacemaker channels activated by hyperpolarization and regulated by cyclic nucleotides. HCN1 and HCN2 play an important role for motor learning and neuronal integration by cerebellar Purkinje cells; as well as, shaping autonomous activity of single neurons and the periodicity of network oscillations. The HCN1 expression is highly enriched in cerebral cortex, hippocampus, cerebellum, and facial motor nucleus. HCN2 is highly abundant in mamillary bodies, pontine nucleus, ventral cochlear nucleus, and nucleus of the trapezoid body. These variations in regional specificity of HCN channels could generate important differences in neuronal pacemaker activity across brain systems. FUNCTION: Hyperpolarization-activated ion channel exhibiting weak selectivity for potassium over sodium ions. Contributes to the native pacemaker currents in heart (If) and in neurons (Ih). Produces a large instantaneous current. Activated by cAMP. Modulated by intracellular chloride ions and pH; acidic pH shifts the activation to more negative voltages. SUBUNIT: The potassium channel is probably composed of a homo- or heterotetrameric complex of pore-forming subunits. Heteromultimer with HCN1. Interacts with KCNE2. SUBCELLULAR LOCATION: Membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Highly expressed in neonatal and adult ventricle and in brain. Highly expressed in the pyramidal layer in hippocampus, in anterior dorsal nucleus in thalamus, in the mammillary nucleus in hypothalamus, in red nucleus, in trigeminal mesencephalic, spinal and principal nuclei, in cochlear and trapezoid nuclei and in the dorsal tegemental nucleus. INDUCTION: By thyroid hormones in hypothyroid animals. DOMAIN: The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position.

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: HCN2