ATP6V1A Overexpression Lysate

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Western Blot: ATP6V1A Overexpression Lysate (Adult Normal) [NBL1-07837] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ATP6V1A.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB
Datasheet
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ATP6V1A Overexpression Lysate Summary

Description

ATP6V1A Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC204425

Accession#: NM_001690

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
ATP6V1A

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
68.1 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 ATP6V1A Overexpression Lysate

  • 70kD, isoform 1
  • ATP6A1
  • ATP6V1A
  • ATP6V1A1
  • ATP6V1A1ATPase, H+ transporting, lysosomal, subunit A1
  • ATPase, H+ transporting, lysosomal (vacuolar proton pump), alpha polypeptide
  • ATPase, H+ transporting, lysosomal 70kDa, V1 subunit A
  • EC 3.6.3
  • EC 3.6.3.14
  • EC 7.1.2.2
  • H(+)-transporting two-sector ATPase, subunit A
  • H+-transporting ATPase chain A, vacuolar (VA68 type)
  • HO68
  • VA68
  • vacuolar ATP synthase catalytic subunit A, ubiquitous isoform
  • Vacuolar ATPase isoform VA68
  • vacuolar proton pump alpha subunit 1
  • Vacuolar proton pump subunit alpha
  • V-ATPase 69 kDa subunit 1
  • V-ATPase 69 kDa subunit
  • V-ATPase A subunit 1
  • V-ATPase subunit A
  • Vma1
  • VPP2
  • V-type proton ATPase catalytic subunit A

Background

This gene encodes a component of vacuolar ATPase (V-ATPase), a multisubunit enzyme that mediates acidification of eukaryotic intracellular organelles. V-ATPase dependent organelle acidification is necessary for such intracellular processes as protein sorting, zymogen activation, receptor-mediated endocytosis, and synaptic vesicle proton gradient generation. V-ATPase is composed of a cytosolic V1 domain and a transmembrane V0 domain. The V1 domain consists of three A and three B subunits, two G subunits plus the C, D, E, F, and H subunits. The V1 domain contains the ATP catalytic site. The V0 domain consists of five different subunits: a, c, c', c', and d. Additional isoforms of many of the V1 and V0 subunit proteins are encoded by multiple genes or alternatively spliced transcript variants. This encoded protein is one of two V1 domain A subunit isoforms and is found in all tissues. Transcript variants derived from alternative polyadenylation exist. [provided by RefSeq]. 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.

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Bioinformatics

Gene Symbol ATP6V1A