LECTIN FROM ARACHIS HYPOGAEA PEROXIDASE LABELED (C005B-455925)

Application

General Western Blot Protocol:

• Glycoprotein sample size: 500ng
• Lectin Concentration: 0.1ug/ml

1. Load samples at 500 ng of glycoprotein per lane
2. Run 4-20% Bis-Tris SDS page gel
3. Transfer gel to a PVDF membrane
4. Block membrane for 1 hr at RT with RIPA buffer (R0278 Sigma)
5. Incubate HRP lectin at 0.1ug/ml with RIPA buffer for 2 hours at RT
6. Wash membrane 5 x 5 minutes with 25ml RIPA buffer
7. 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.

Unit Definition

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

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.