His Detect Solution

For in vitro use only!

Shipping: shipped at ambient temperature

Storage Conditions: store at ambient temperature

Shelf Life: 12 months (3 months upon opening)

Description:
His Detect Solution is a ready-to-use reagent for detection of poly-histidine tagged proteins in blot assays. The reagent is equally suitable for Western Blot, Dot or Slot Blot assays. The detection reagent is made from an optimized anti-histidine antibody gold nanoparticle conjugate and is reactive against C- and anti-N terminal tags. The protein of interest is specifically stained in pink directly on the blot. Instant detection of poly-histidine tagged proteins on nitrocellulose membrane with His Detect is easy and does not require relevant hands on time. No blocking, Washing or successive antibody incubation time needed. A detection limit of 0.1 pmol can be achieved with this highly sensitive detection reagent. This is in the range of conventional ECL detection.

Content:
30 ml His Detection reagent

Additional Materials Required:

• Incubation tray
• Lab shaker

Preparation Procedure:

• Electro blotting is performed as usual, e.g. protein from SDS-PAGE is transferred onto Nitrocellulose (NC) or Polyvinylidenfluorid (PVDF) membrane. Alternatively, fractions from cell extracts or column purifications are spotted onto a suitable membrane. In any case the membrane should be quickly rinsed with plain tap water to remove remaining detergents.
• The membrane is placed into a clean incubation tray of suitable size. 10 ml of His Detect ready-to-use reagent is poured onto the membrane to cover it completely. Incubation is performed on a suitable lab shaker. In approx. one hour His-tagged fusion proteins become visible due to antibody induced gold nanoparticle accumulation.
• After reaching a sufficient staining intensity the excess reagent should be removed and collected in a separate bottle. Depending on the number of protein spots, the His Detection reagent may be re-used for up to two more staining procedures.
• The membrane is rinsed with water and can be dried with filter paper and archived or scanned.

There is no detection equipment required. The detection signal is visible by the human eye and may be quantified by spectroscopic or photo-technical equipment. The intensity of the band is proportional to the amount of protein present, thus quantification can be easily achieved with standard spotted next to the sample.