I have phosphate in my enzyme. What can I do?

You can dialyse or desalt the enzyme into a phosphate-free buffer. Alternatively, you can use a special resin (PiBind) to remove the phosphate.

I have 5% DMSO in my assay. Can I use PiColorLock Gold?

Yes, the reagent is designed for drug screening work and other situations that require DMSO.

I have a high background in my ATPase assay and I definitely do not have free phosphate in my sample

This is almost always due to inadequate mixing of the special stabilizer with the sample and detection reagent. Make sure the stabilizer is pipetted up and down several times to ensure thorough mixing.

I was wondering if it is possible to buy the purified ATP that comes in the following kit separately? We bought the whole kit earlier, but we are in need for more of the purified ATP component from the kit.

We do have extra ATP from the kit that we can send you no problem. Please send me you original order information and I can use that to get everything processed and sent out to the correct location.

I would like to measure the conversion of pyrophosphate to phosphate. Can I use the PiColorLock Gold Phosphate Detection System for this purpose?

Yes, only the phosphate will give a signal; pyrophosphate will not.

I would like to work with ecto-ntpdase enzymes in human plasma samples. What kind of samples can it be used with the ATPase assay kit?

In principle our ATPase assay kits can be used to measure any ATPase. With complex samples however (e.g. membranes, homogenates) you will need to be able to distinguish hydrolysis of ATP mediated by the enzyme of interest, and hydrolysis mediated by other ATPases. Unfortunately, we do not know whether other enzymes may be present. If other enzymes are present, you will need to find assay conditions in which your enzyme is the dominant activity (by manipulating pH, buffer composition or perhaps through using specific inhibitors of the other enzymes). Another approach that some people use is to immunoprecipitate the ATPase of interest before the assay is carried out, which effectively removes all other enzymes.

I'd like to detect Na-K ATPase activity. Can the product Colorimetric ATPase Assay Kit (601-0120) detect specific Na-K ATPase activity?

This kit is intended for the measurement of ATPase activity, but is not specific to one particular type of activity.

ATPase Activity Kit (Colorimetric)

Name: ATPase Activity Kit (Colorimetric)
Brand: Novus Biologicals
SKU: 601-0120
Rating: 4 (1 reviews)

Novus Biologicals, part of Bio-Techne | Catalog # 601-0120

(1)

Citations (71)

Product Datasheet / COA / SDS / Protocols & Manuals

Discontinued Product

601-0120 has been discontinued. View all ATPase products.

Product Specifications
Kit Contents
Preparation & Storage
Background
Product Documents
Specific Notices

Product Specifications

Kit Type

Activity Kit (Colorimetric)

Specificity

Na+/K+-ATPase activity determination (PMID: 21304972)

Description

This non radioactive colorimetric assay kit contains all the necessary reagents for measuring enzyme activity (everything included in the PiColorlock kit and more) and is ideal for high throughput drug screening.

The kits contain PiColorlock, a non-radioactive, superior phosphate detection reagent.

Key Features of PiColorLock:

Colorimetric Assay-competitor assays are radioactive.
Special additive ensures low backgrounds with acid-labile substrates.
Unparalleled stability of phosphate-dye complexes.
Reagent is compatible with almost any assay buffer.
No inhibition of color development by high concentrations of protein.
Stable reagent formulation - long shelf life.

Colorimetric assays for ATPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions.

Kit Contents for ATPase Activity Kit (Colorimetric)

1 x 0.5ml of Accelerator ( 1 x 0.5ml)
1 x 1.5 ml of 0.1M MgCl2 (2 x 1.5 ml)
1 x 10 ml of PiColorLock ( 1 x 10ml)
1 x 5 ml of 0.1mM Pi standard (1 x 5ml)
1 x 5ml of 0.5M Tris PH 7.5 (1x 5ml)
1 x 5ml of Stabilizer (1 x 5ml)
2 x 96-well plates
4 vials of lyophilized ATP (4 vials)

Preparation and Storage

Formulation

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage of components varies. See protocol for specific instructions.

Background: ATPase

CATALYTIC ACTIVITY: ATP + H2O = ADP + phosphate. SUBCELLULAR LOCATION: Membrane; multi-pass membrane protein (By similarity). ALTERNATIVE PRODUCTS: 2 named isoforms produced by alternative splicing. SIMILARITY: Belongs to the cation transport ATPase (P-type) family. Type V subfamily.

Alternate Names

ATPase, axonemal, dynein, axonemal, heavy chain 8, dynein, axonemal, heavy polypeptide 8, FLJ25850, FLJ36115, FLJ36334, HDHC9

Gene Symbol

DNAH8

Additional ATPase Products

Product Documents for ATPase Activity Kit (Colorimetric)

Product Datasheets

Download Product Datasheets

Protocols & Manuals

Download Manual - ATPase Activity Kit (Colorimetric)

COA

SDS

Download SDS

Product Specific Notices for ATPase Activity Kit (Colorimetric)

This product is manufactured by Abcam and distributed by Novus Biologicals.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. This product is guaranteed for 1 year from date of receipt and this statement overrides any mentioned guarantee period on the limitations section of this products datasheet. Please contact technical@novusbio.com with questions.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Citations
Reviews