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TRPV6 Overexpression Lysate

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Western Blot: TRPV6 Overexpression Lysate (Adult Normal) [NBL1-17345] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TRPV6.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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TRPV6 Overexpression Lysate Summary

Description

TRPV6 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC214982

Accession#: NM_018646

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
TRPV6

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
83 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 TRPV6 Overexpression Lysate

  • ABP/ZF
  • calcium channel CaT1
  • Calcium transport protein 1
  • CaT1
  • CATL
  • CaT-L
  • CaT-like
  • ECAC2
  • ECAC2caT-L
  • epithelial apical membrane calcium transporter/channel CaT1
  • Epithelial calcium channel 2Alu-binding protein with zinc finger domain
  • HSA277909
  • LP6728
  • transient receptor potential cation channel subfamily V member 6
  • transient receptor potential cation channel, subfamily V, member 6
  • TRPV6
  • ZFAB

Background

Calcium-permeable channels, such as TRPV6, participate in neurotransmission, muscle contraction, and exocytosis by providing calcium as an intracellular second messenger. Depending on the tissue, transcellular calcium transport may be regulated by vitamin D, parathyroid hormone (PTH), or calcitonin (CALCA); FUNCTION: Calcium selective cation channel probably involved in Ca(2+) uptake in various tissues, including Ca(2+) reabsorption in intestine. The channel is activated by low internal calcium level, probably including intracellular calcium store depletion, and the current exhibits an inward rectification. Inactivation includes both, a rapid Ca(2+)-dependent and a slower Ca(2+)-calmodulin-dependent mechanism, the latter may be regulated by phosphorylation. In vitro, is slowly inhibited by Mg(2+) in a voltage-independent manner. Heteromeric assembly with TRPV5 seems to modify channel properties. TRPV5-TRPV6 heteromultimeric concatemers exhibit voltage-dependent gating. SUBUNIT: Homotetramer and probably heterotetramer with TRPV5. Interacts with TRPV5. Interacts with S100A10 and probably with the ANAX2-S100A10 heterotetramer. The interaction with S100A10 is required for the trafficking to the plasma membrane. Interacts with BSPRY. Interacts with calmodulin. SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.

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.

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Bioinformatics

Gene Symbol TRPV6