Talk:Immunolabeling

Comments from MartinLubell about Immunolabeling Wikipedia article

I think that overall your article was very informative. I definitely learnt a lot about "immunolabeling"! It's very cool that you were able to get permission from Dr. Thomas Caceci to use his images using the "immunolabeling" technique. I can't wait to see them.

Following are a few small edits:

• Issues with singular and plurals: use "piped links" to solve problem.

Antigens are organic molecules, proteins, capable of binding to antibodies. Linking antigen-specific antibodies with a means of detection, known as a tag, provides a way to observe the location of the antigens to which the antibodies bind. If the immunolabeling process is meant to reveal information about a cell or its substructures, the process is called immunocytochemistry. Immunolabeling of larger structures, such as biological tissues, is called immunohistochemistry.

There are two complex steps in the manufacture of antibodies for immunolabeling. The first is the production of antibodies that bind specifically to the antigen of interest and the second is the fusion of the tag to the said antibodies. Since it is impractical to fuse a tag to every conceivable antigen-specific antibody, most immunolabeling processes use an indirect method of detection. This indirect method employs a primary antibody that is antigen-specific and a secondary antibody fused to a tag that specifically binds the primary antibody. This indirect approach permits mass production of secondary antibodies that can be bought off the shelf.[1]

• 3rd paragraph: Add enzyme link
• 4th paragraph: question to myself: if the gold is so dense that it is impenetrable, why is there radiation from it? Wouldn't it be more of a black spot?
• Very good 5th paragraph!
• 8th paragraph: extra space at end of: "... haptenylated primary antibodies can be used ,"
• 9th paragraph: missing some links for "tag" & "antibodies"
• 10th paragraph: For references, should there be a space before the number or not? (I'm not sure.)

MartinLubell (talk) 06:17, 11 April 2013 (UTC)

Martin, thank you for all your editorial advice. I went ahead and corrected the spacing in the 8th paragraph. Good eye! I think you are right on the spacing before the number of the references. Those have been corrected. My partner and I will definitely consider your other suggestions as we continue to work on this article over the next few weeks. Thanks, again! Jbmontgomery24 (talk) 12:22, 11 April 2013 (UTC)

You are very welcome! Ciao, MartinLubell (talk) 12:37, 11 April 2013 (UTC)

Primary antibody tagging

This article only says that the secondary antibody can be tagged, but the article Direct fluorescent antibody indicates that the primary antibody can sometimes be tagged. M-1 (talk) 11:03, 14 April 2012 (UTC)

Correct. Secondary antibody tagging has the major advantage of cost, the way it's usually done. One can produce a "normal" (untagged; primary) antibody for the specific antigen of interest, then use a mass-produced tagged secondary antibody to locate it. Otherwise, one would have to not only get the initial primary antibody, but do the work to get it properly tagged for localization, which would likely cost more and take more time. I'll try to locate some references on this that are accessible to others... Allens (talk | contribs) 22:14, 27 May 2012 (UTC)

Improvements in Need for the Immunolabeling Article

Greetings, fellow Wikipedians! This article needs to be greatly improved in the following ways:

• Add verified references and inline citations within the lead section
• Add free copyright images throughout the article for a better visual understanding of this technique
• Provide some general statements (with references) within the lead section for any reader to have a better grasp of its importance
• Provide subsections after the lead section
• Overview of antibodies and their role in immunolabeling
• Provide examples where immunolabeling is used
• References, references, and more references
• If feasible, find a free copyright video of the technique being used

Jbmontgomery24 (talk) 00:27, 13 March 2013 (UTC)

Comments from Tisquestra about article improvement

This article has been greatly modified, and contains a lot of well curated information. A couple things for further improvement..

In terms of formatting and content:

• The table of contents should probably be at the top, perhaps next to the introductory paragraph
• The description of the antibody experiment (2nd, 3rd, and 4th paragraph) contains a rather basic image, that although serves as a visual of what is being described, may not be properly detailed to the user. Perhaps a visual of an tagged antibody, that shows the user how a researcher can visualize the second antibody in the test system.
• The last full paragraph in the introductory section should perhaps belong in its own section. The paragraph starts with describing the applications of immunolabeling, but only one application (identifying a drug on a particular organ) is described for the rest of the paragraph.
• Because the two sections "Immunolabeling for Electron Microscopy" and "Immunolabeling for Scanning Electron Microscopy" are so similar, it is probably a good idea to delineate the differences between these two techniques more clearly. You compare the scanning method to the fluorescence method, so the technique is indeed different, but (maybe for myself) why are the names so similar? What is the similarity between them?

Citation-wise:

• The first paragraph, although a good introduction, does not provide any citations.
• Although your second paragraph does have a citation, it refers to a book which is not accessible readily online. Perhaps, additionally, one could use an online resource as well to back up the experiment described?
• In your section "Indirect vs. Direct Methods" it is not clear how the citation in the first paragraph gives the information that you provided. Rather, I see it as being more useful to describe the second paragraph only.

Suggestions for Lead Section Structure

Since this article is being created within the context of already existent articles on immunocytochemistry and immunohistochemistry, I recommend that the focus of this article should be on the purposes of immunolabeling in general and the fundamental tools: Ab, tag, signal resolution, etc. I recommend avoiding any detailed discussion of the procedures that would overlap with the articles mentioned.

My initial thoughts on the structure of the lead are as follows:

1.) I like the first sentence. I suggest the following sentence indentify, distinguish and link to the immunocytochemistry and immunohistochemistry subsets. This will signal to the reader that immunolabeling is a very broad procedure; the essence of which involves just a few elements.

2.) Explain the type of information that can be obtained through immunolabeling.

3.) Identify the elements and distinguish between the direct and indirect approach.

4.) Explain why the indirect approach is more practical.

5.) Briefly describe the binding dynamics of antibody and how they are produced.

6.) Briefly describe some common tags and how they are fused to secondary antibody.

7.) Discuss the resolution of the tag signal.Jakodak (talk) 13:08, 13 March 2013 (UTC)

Lead Section Edits

Hello all! I currently have the opportunity to help with this article's status through a Molecular Biology course that I am taking through a graduate program at JHU. For this project, my partner (Jakodak) and I will do our best to continue to develop this article. A lot of editing will be necessary in the lead section due to the lack of references. Anyone oppose to the lead section being redone immensely due to no sources in this section? If not, we will start our new additions and edits shortly. Thanks! Jbmontgomery24 (talk) 22:38, 31 March 2013 (UTC)