Q: We have received an inquiry from our customer who is interested in your product, ATP6V1B1 Overexpression Lysate (Catalog#NBL1-07838). Our customer would like to know a detail information about Western Blot image(an attached image) on your website. Could you tell us information (Catalog Number, etc.) about the antibody stained the band by Western Blot ?

A: The Western blot image shown on the webpage for our ATP6V1B1 over-expression lysate with catalogue number NBL1-07838 was generated using an empty vector transfected control cell lysate (HEK293 cell lysate), and an over-expression lysate for ATP6V1B1. The ATP6V1B1 has a C-terminal Myc/DDK (Flag) tag and an anti-DDK antibody was used for detection. The predicted molecular weight of ATP6V1B1 is 56.8kDa. Here, for your reference, is the Uniprot listing for this protein: UniProt P15313.

ATP6V1B1 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-07838

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for ATP6V1B1 Overexpression Lysate

ATP6V1B1 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC209462

Accession#: NM_001692

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.

Product Specifications

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

56.7 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.

Type

Overexpression

Scientific Data Images for ATP6V1B1 Overexpression Lysate

Western Blot: ATP6V1B1 Overexpression Lysate [NBL1-07838]

Western Blot: ATP6V1B1 Overexpression Lysate (Adult Normal) [NBL1-07838] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ATP6V1B1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: ATP6V1B1

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 B subunit isoforms and is found in the kidney. Mutations in this gene cause distal renal tubular acidosis associated with sensorineural deafness. [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.

Alternate Names

ATP6B158kD subunit, ATPase, H+ transporting, lysosomal 56/58kDa, V1 subunit B1, Endomembrane proton pump 58 kDa subunit, H+-ATPase beta 1 subunit, vacuolar proton pump 3, Vacuolar proton pump subunit B 1, vacuolar proton pump, subunit 3, VATBMGC32642, V-ATPase B1 subunit, V-ATPase subunit B 1, Vma2, V-type proton ATPase subunit B, kidney isoform

Gene Symbol

ATP6V1B1

Additional ATP6V1B1 Products

Product Documents for ATP6V1B1 Overexpression Lysate

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Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for ATP6V1B1 Overexpression Lysate

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.

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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