ab Optiblot Optiblot Luminol Membrane Pen

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1 ab Optiblot Optiblot Luminol Membrane Pen Instructions for Use For use in Western Blot. This product is for research use only and is not intended for diagnostic use.

2 Table of Contents 1. Introduction 2 2. Kit Components 3 3. Storage 3 4. Using the Optiblot Luminol Membrane Pen 4 1

3 1. Introduction Western blotting (WB) is a key technique in current life science research. By utilizing monoclonal or polyclonal antibodies to specific antigen proteins, it is possible to identify the signal of a protein of interest, quantitatively or qualitatively, on blotting membranes. However, due to the complexities of biological materials and the insufficient specificity of some utilized antibodies, some intriguing WB results are generally observed, such as multiple band signals or hybridized bands with unclear molecular weights. To validate a WB result, calculating the molecular weight of the obtained protein signals may be the most straight forward means. Nevertheless, since most (pre-stained) molecular weight standard markers cannot be visualized after the hybridization procedure, many laboratories may align the WB result with the original membrane blot. Such procedure is tedious and may introduce estimation bias. The Optiblot Luminol Membrane Pen is designed to simplify such tasks and improve accuracy. After blotting proteins from gels to PVDF or nitrocellulose membranes, just use the Optiblot Luminol Membrane Pen to mark the (pre-stained) molecular weight standard markers and perform regular hybridization procedures. During the colorimetric or chemiluminenscent development step, the bands of molecular weight standard marker can be visualized and recorded for further analysis. Susceptible protein bands are easily located 2

4 from the WB result. (red arrow indicated). Additionally, in WB application, the Optiblot Luminol Membrane Pen can be used for blot annotation. 2. Kit Components Constituent Quantity Optiblot Luminol Membrane Pen 1 3. Storage The Optiblot Luminol Membrane Pen should be stored at +4 C. It should not be frozen. The strength of drawing signal will gradually decrease six months after the first usage. 3

5 4. Using the Optiblot Luminol Membrane Pen 1. During electrophoresis, run pre-stained molecular weight standard markers in parallel with the samples. 2. After electrophoresis and membrane blotting procedures, gently remove the residual solution from the transferred PVDF or nitrocellulose membranes by using filter paper. Keep the membranes wet and do not let the membranes over-dry. 3. Mark the visible pre-stained protein bands evenly with the Optiblot Luminol Membrane Pen. Generally, one drawing should be enough for delivering strong signals. If necessary, repeatedly marking the same location for stronger signals. The Optiblot Luminol Membrane Pen can also be used for annotation on the margin of membrane. 4. Perform membrane blocking and the rest of all necessary procedures for western blotting. No particular care has to be taken. 4

6 5. Troubleshooting Problem Reason Solution No marked signal/ marked signal too weak Optiblot Luminol Membrane Pen has expired The strength of drawing signal will gradually decrease six months after the first usage. Please use an un-expired Optiblot Luminol Membrane Pen Less effective developing system (e.g. colorimetric system) or substrate was employed Apply more than one drawing on the target area. The marked signal is too strong Too effective developing substrate was employed. Use a less effective developing substrate. 5

7 For hints and tips on Western Blotting as well as other techniques please visit Keep up to date on Abcam events and conferences on Facebook & Twitter. 6

8 UK, EU and ROW Tel: +44-(0) Austria Tel: France Tel: Germany Tel: Spain Tel: Switzerland Tel (Deutsch): Tel (Français): US and Latin America Tel: ABCAM (22226) Canada Tel: China and Asia Pacific Tel: ( 中國聯通 ) Japan technical@abcam.co.jp Tel: +81-(0) Copyright 2013 Abcam, All Rights Reserved. The Abcam logo is a registered trademark. All information / detail is correct at time of going to print.