LowCross-Buffer®

LowCross-Buffer® is not for use as a blocking solution for surfaces!

Product description

pH-value: pH 7.2 ± 0.2
Preservative: contains 0.1% ProClin® 300
Storage: shelf life at –20 °C: 1 year – tolerates repeated freezing and thawing cycles
shelf life at 2 – 8 °C: 6 months

For research use only, not for diagnostic use

Available package sizes:

50   ml order number 100 050
125 ml order number 100 125
500 ml order number 100 500

Instructions for use

The newly developed LowCross-Buffer® lowers cross reactivities, nonspecific binding and matrix effects in immunoassays like ELISA, EIA, Western blotting, immuno-PCR, protein arrays, multianalyte immunoassays and immunohistochemistry – depending on the characteristics of the assay type and the used antibodies.

Immediately before use the buffer should be mixed thoroughly. LowCross-Buffer® is used instead of a sample buffer or antibody dilution buffer for the immunological reaction.

LowCross-Buffer® is unsuitable for blocking of surfaces. For blocking of surfaces we recommend The Blocking Solution (Order No. 110 050, 110 125 and 110 500). LowCross-Buffer® is not useable as a sample buffer for electrophoresis.

Examples of use:
ELISA: dilution buffer for specimen and for the detection antibodies
Western blotting: dilution buffer for primary and secondary antibodies
Immunohistochemistry: dilution buffer for primary and secondary antibodies
Protein arrays: dilution buffer for specimen and for the detection antibodies

Dilution of the specimen:
Standards and specimen for ELISA and protein arrays can be diluted with LowCross-Buffer® at 1:2 or higher. Standards and specimen should be treated strictly the same way.

Dilution of antibodies: Antibodies can be diluted with LowCross-Buffer® in a user-defined manner, depending on the recommendation of the data sheet of the antibodies. This is the same for primary and secondary antibodies.

Appearance of signal reduction:
In some cases a smooth reduction of the wanted signal can be observed. LowCross-Buffer® reduces low- and middle-affinity binding. That means that by the use of low- and middle-affinity antibodies or polyclonal antibodies a smooth reduction of signals can appear. Polyclonal antibodies normally contain low- and middle-affinity binding components.

In the case of polyclonal antibodies a moderate increase of the concentration of the antibody can lead to the previously seen signals. Unwanted low and middle-affinity binding will be still reduced by LowCross-Buffer®. In the case of low- and middle affinity antibodies (also monoclonal antibodies) a pre-dilution of LowCross-Buffer® with salt-free water can be useful to get the previously seen signal. But in this case also the unwanted bindings or cross-reactivities can partly occur again, depending on the choosen dilution with water.

Although LowCross-Buffer® is used as an assay buffer it is necessary to saturate surfaces like ELISA-wells or membranes with a blocker. We recommend the use of The Blocking Solution (Order No. 110 050, 110 125 and 110 500).

LowCross-Buffer® can be used also as washing buffer – especially in delicate or interference-sensitive assays like immuno-PCR.

Components of immunoassays – as well as possibly of LowCross-Buffer® – maybe quench the fluorescence of fluorescein dyes. Therefore we recommend the use of Oyster*- (Denovo Biolabels), CyDye**- (Amersham) or Alexa***- (Molecular Probes) fluorescence dyes.

We strongly recommend that the effectiveness of LowCross-Buffer® for a certain application is tested before.

*Oyster is a registered trade mark of the company Denovo Biolabels.
**CyDye is a registered trade mark of the company Amersham Biosciences.
***Alexa Fluor Dye is a registered trade mark of the company Molecular Probes.

LowCross-Buffer is a registered trade mark of CANDOR Bioscience GmbH.
ProClin is a registered trade mark of Rohm and Haas Company.

For challenging applications we have developed two additional formulations of LowCross-Buffer® called LowCross-Buffer® MILD and LowCross-Buffer® STRONG.