Application

General Western Blot Protocol: Glycoprotein sample size: 500ng Lectin Concentration: 0.1ug/mlLoad samples at 500 ng of glycoprotein per lane Run 4-20% Bis-Tris SDS page gel Transfer gel to a PVDF membrane Block membrane for 1 hr at RT with RIPA buffer (R0278 Sigma) Incubate HRP lectin at 0.1ug/ml with RIPA buffer for 2 hours at RT Wash membrane 5 x 5 minutes with 25ml RIPA buffer Detect using chemiluminescent substrate (CPS1-120)

Biochem/physiol Actions

PNA does not agglutinate normal human erythrocytes, but strongly agglutinates neuraminidase treated erythrocytes. PNA has potent anti-T activity similar to the anti-T antibody in human sera. The lectin can be used to distinguish between human lymphocyte subsets.

Packaging

250 µg in serum bottle

Physical form

Contains sodium citrate

Preparation Note

Prepared from peroxidase (P8375) using a modification of a published method, which favors low molecular weight conjugates. Repurified after conjugation by affinity chromatography.

Unit Definition

One unit will form 1 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.