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	<id>https://wiki.esonto.de/index.php?action=history&amp;feed=atom&amp;title=Immunofluorescence_staining</id>
	<title>Immunofluorescence staining - Revision history</title>
	<link rel="self" type="application/atom+xml" href="https://wiki.esonto.de/index.php?action=history&amp;feed=atom&amp;title=Immunofluorescence_staining"/>
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	<updated>2026-04-17T19:26:23Z</updated>
	<subtitle>Revision history for this page on the wiki</subtitle>
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	<entry>
		<id>https://wiki.esonto.de/index.php?title=Immunofluorescence_staining&amp;diff=129&amp;oldid=prev</id>
		<title>ESO wikiadmin: Created page with &quot;{| class=&quot;wikitable&quot; |+ !Materials ! ! ! |- | |PBS++ (50 ml PBS; 25 μl 2.5M CaCl&lt;sub&gt;2&lt;/sub&gt; ; 1M 50 μl MgCl&lt;sub&gt;2&lt;/sub&gt;) | | |- | |Permeabilization solution (PBS++; 10% FCS; 0.2% Saponin; alternatively 0.2% | | |- | |Triton X-100 may be used instead of saponin as a detergent) | | |- | |Blocking solution (PBS++; 10% FCS) | | |- | |First and second antibody | | |- | |Mounting medium | | |- | |Nail polish | | |}  === Procedure === ----  * After fixation wash (3 x with PB...&quot;</title>
		<link rel="alternate" type="text/html" href="https://wiki.esonto.de/index.php?title=Immunofluorescence_staining&amp;diff=129&amp;oldid=prev"/>
		<updated>2025-03-07T09:47:55Z</updated>

		<summary type="html">&lt;p&gt;Created page with &amp;quot;{| class=&amp;quot;wikitable&amp;quot; |+ !Materials ! ! ! |- | |PBS++ (50 ml PBS; 25 μl 2.5M CaCl&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; ; 1M 50 μl MgCl&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;) | | |- | |Permeabilization solution (PBS++; 10% FCS; 0.2% Saponin; alternatively 0.2% | | |- | |Triton X-100 may be used instead of saponin as a detergent) | | |- | |Blocking solution (PBS++; 10% FCS) | | |- | |First and second antibody | | |- | |Mounting medium | | |- | |Nail polish | | |}  === Procedure === ----  * After fixation wash (3 x with PB...&amp;quot;&lt;/p&gt;
&lt;p&gt;&lt;b&gt;New page&lt;/b&gt;&lt;/p&gt;&lt;div&gt;{| class=&amp;quot;wikitable&amp;quot;&lt;br /&gt;
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!Materials&lt;br /&gt;
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|PBS++ (50 ml PBS; 25 μl 2.5M CaCl&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; ; 1M 50 μl MgCl&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;)&lt;br /&gt;
|&lt;br /&gt;
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|Permeabilization solution (PBS++; 10% FCS; 0.2% Saponin; alternatively 0.2%&lt;br /&gt;
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|-&lt;br /&gt;
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|Triton X-100 may be used instead of saponin as a detergent)&lt;br /&gt;
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|Blocking solution (PBS++; 10% FCS)&lt;br /&gt;
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|-&lt;br /&gt;
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|First and second antibody&lt;br /&gt;
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|Mounting medium&lt;br /&gt;
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|Nail polish&lt;br /&gt;
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|}&lt;br /&gt;
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=== Procedure ===&lt;br /&gt;
----&lt;br /&gt;
&lt;br /&gt;
* After fixation wash (3 x with PBS++) the cover slips in the 24 well/plates&lt;br /&gt;
* Add 300 μl/well permeabilization solution (10 min)&lt;br /&gt;
* Wash samples 3 x with PBS++&lt;br /&gt;
* Incubate samples with 300 μl/well blocking solution (5 min)&lt;br /&gt;
* Add dropwise 25 μl/well of the first antibody (diluted 1:100 to 1:400 in blocking solution) on the cover slip&lt;br /&gt;
* Inubate 1 h at RT&lt;br /&gt;
* Wash samples 3 x with PBS++&lt;br /&gt;
* Incubate samples with 300 μl/well blocking solution (5 min)&lt;br /&gt;
* Add on drops 25 μl/well of the fluorescent coupled second antibody (diluted 1:100 to 1:200 in blocking solution) on the cover slip&lt;br /&gt;
* Incubate for 45 min at RT &amp;#039;&amp;#039;&amp;#039;in the dark&amp;#039;&amp;#039;&amp;#039;&lt;br /&gt;
* Wash samples 3 x with PBS++&lt;br /&gt;
* Transfer cover slips on the glass slide (add mounting medium in between)&lt;br /&gt;
* After drying seal the cover slips with nail polish.&lt;/div&gt;</summary>
		<author><name>ESO wikiadmin</name></author>
	</entry>
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