One of our customer ordered antibody to aquaporin 3 cat# NBP1-46502. She doesn't manage to make it work in WB so she wanted to know if you can advise on what molecular might she can except to see the band? She also wanted to order the positive control lysate NBL1-07638 but there are so many bands on the blot that she's afraid it won't help! Could you tell me which antibody was used on the blot picture we can see on the web? And what dilution was used? Could you just confirm if the image from NBL1-07638 datasheet was obtained with antibody NBP1-46502 or another antibody? Then as there are so many bands, would you be able to supply a blocking peptide to switch off the specific band? Which kind of washing buffer and blocking buffer do you suggest? Which steps (how many and how long would you suggest)?

Name: Aquaporin-3 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-07638

The Theoretical MW of AQP3 is 32 KDa. This antibody is unpurified and therefore may detect more unspecific bands than a purified antibody. I would recommend increasing the blocking and washing times to alleviate this issue, along with diluting the antibody further. NBP1-64502 was not used on NBL1-07638, and while it should work I would recommend trying the suggestions above first. Unfortunately we don't have a blocking peptide available for this product.

Aquaporin-3 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-07638

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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 Aquaporin-3 Overexpression Lysate

Aquaporin-3 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC201856

Accession#: NM_004925

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.

TMW

31.4 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 Aquaporin-3 Overexpression Lysate

Western Blot: Aquaporin-3 Overexpression Lysate [NBL1-07638]

Western Blot: Aquaporin-3 Overexpression Lysate (Adult Normal) [NBL1-07638] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Aquaporin-3.

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: Aquaporin-3

Aquaporin 3 is a water channel protein. Aquaporins are a family of small integral membrane proteins related to the major intrinsic protein (MIP or AQP0). Aquaporin 3 is localized at the basal lateral membranes of collecting duct cells in the kidney. In addition to its water channel function, aquaporin 3 has been found to facilitate the transport of nonionic small solutes such as urea and glycerol, but to a smaller degree. It has been suggested that water channels can be functionally heterogeneous and possess water and solute permeation mechanisms. [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

AQP-3, Aquaglyceroporin-3, aquaporin 3, aquaporin 3 (GIL blood group), aquaporin 3 (Gill blood group), aquaporin-3, GIL, Gill blood group

Gene Symbol

AQP3

Additional Aquaporin-3 Products

Product Documents for Aquaporin-3 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Aquaporin-3 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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