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  • In this final phase, we will demonstrate signal development using the most common,

  • most sensitive and most inexpensive detection method the

  • electrochemiluminescence (or ECL) reaction.

  • This method utilizes the HRP enzyme,

  • which was conjugated to the secondary

  • to catalyze the ECL reaction and produce light.

  • A light is then gathered onto x-ray film and developed or digitized with the

  • aid of a specialized camera sensitive enough for this application.

  • We start by mixing equal parts ECL reagents in a one-to-one ratio

  • according to the manufacturer's instructions.

  • We will incubate the membrane for 3-5 minutes without agitation.

  • After incubation, decant ECL mixture and use a laboratory wipe to wipe off excess

  • solution

  • from the corner of the membrane.

  • Place the membrane in a clear plastic wrap such as a sheet protector to

  • prevent drying.

  • Avoid letting the membrane completely dry out.

  • We can now use a roller to push out any bubbles or any excess solution.

  • Immediately develop the membrane.

  • Both film and camera systems allow you to manually adjust the exposure time

  • in order to ensure a picture perfect Western Blot. Relative band densities can

  • now be quantified with commercially available software.

  • Proper molecular weight can also be verified by comparing band sizes to the

  • molecular weight ladder.

In this final phase, we will demonstrate signal development using the most common,

Subtitles and vocabulary

B2 membrane blot detection phase molecular protocol

Western Blot Visual Protocol: Phase 5: Detection

  • 30 3
    Yu Sheng Chen posted on 2016/11/26
Video vocabulary

Keywords

demonstrate

US /ˈdɛmənˌstret/

UK /'demənstreɪt/

  • other
  • To explain or describe something clearly.
  • To show something clearly by giving proof or evidence.
  • verb
  • To display a feeling or ability openly
  • To protest about something often as a group
  • To prove or make clear by reasoning or evidence
  • To show how something works (e.g. product)
  • other
  • To take part in a public demonstration or protest.
completely

US /kəmˈpliːtli/

UK /kəmˈpli:tli/

  • adverb
  • In every way or as much as possible; totally.
  • In every way or as much as possible
  • Totally; entirely.
  • To the greatest extent; thoroughly.
  • In every way or as much as possible; totally.
  • Including all or everything; without anything lacking.
  • Thoroughly; to a full or finished extent.
  • Totally; in every way or as much as possible.
immediately

US /ɪˈmidiɪtli/

UK /ɪˈmi:diətli/

  • adverb
  • Without any delay; straight away
  • Without any delay; at once.
  • Very closely in space or time; next to.
  • Directly and personally.
  • Following or preceding without any interval of time or space.
  • Without any delay; instantly.
common

US /ˈkɑmən/

UK /'kɒmən/

  • noun
  • Area in a city or town that is open to everyone
  • A piece of open land for public use.
  • A piece of open land for public use.
  • Field near a village owned by the local community
  • adjective
  • Lacking refinement; vulgar.
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  • (of a noun) denoting a class of objects or a concept as opposed to a particular individual.
  • Without special rank or position; ordinary.
  • Shared; Belonging to or used by everyone
  • Typical, normal; not unusual
  • Lacking refinement; vulgar.
  • Found all over the place.
wrap

US /ræp/

UK /ræp/

  • noun
  • Piece of clothing worn loosely on the shoulders
  • Thin bread filled with food and rolled
  • verb
  • To be or move around something
  • To cover something with paper or a cloth e.g. gift
method

US /ˈmɛθəd/

UK /'meθəd/

  • noun
  • (Organized and planned) way of doing something
  • A particular form of procedure for accomplishing or approaching something, especially a systematic or established one.
  • A procedure associated with an object class.
  • Orderliness of thought, arrangement, or behavior.
  • A systematic or established way of doing something.
  • other
  • Orderliness of thought, arrangement, or behavior.
proper

US /ˈprɑpɚ/

UK /'prɒpə(r)/

  • adjective
  • Correct according to social or moral rules
  • Correct or appropriate; suitable for the purpose or occasion.
  • Conforming to conventionally accepted standards of behavior or morals.
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  • (of a noun) denoting a particular person, place, or thing.
  • Thorough; complete.
  • Behaving in a polite and socially acceptable way.
develop

US /dɪˈvɛləp/

UK /dɪ'veləp/

  • verb
  • To explain something in steps and in detail
  • To create or think of something
  • To grow bigger, more complex, or more advanced
  • To make a photograph from film
  • other
  • To invent something or cause something to exist
  • To start to suffer from an illness or other medical condition
  • To improve the quality, strength, or usefulness of something
  • other
  • To (cause something to) grow or change into a more advanced, larger, or stronger form
exposure

US /ɪkˈspoʒɚ/

UK /ɪk'spəʊʒə(r)/

  • noun
  • Allowing light through a cameras lens onto film
  • Being talked about in the media
  • Act of making something that is secret known
  • Experience of something directly
  • other
  • The state of being exposed to something; the act of revealing something, especially something scandalous or previously hidden.
  • The state of being unprotected from something harmful, such as the weather.
  • The total amount that someone could lose in an investment or business deal.
  • The condition of being subjected to extreme weather conditions, especially cold, leading to hypothermia.
  • The amount of light allowed to reach a photographic film or sensor.
  • Publicity; the state of being known or recognized.
  • The experience of being exposed to something, such as an idea or culture.
prevent

US /prɪˈvɛnt/

UK /prɪ'vent/

  • verb
  • To stop something from happening or existing
  • other
  • To stop something from happening or someone from doing something.

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