4 Steps to Better Immunohistochemistry: Steps 2, 3, and 4


Posted by Carolyn P on May 31, 2017 3:00:00 AM

This is part two of a two-part series on how to optimize your IHC protocols. Part one introduced the principles behind antigen retrival. Click here if you missed it... but, if you've got your tissue prepped and ready to go, we'll move on to the next steps in the staining protocol.

READ MORE >

Topics: Protocols, IHC, techniques

4 Steps to Better Immunohistochemistry: Step 1 - Antigen Retrieval


Posted by Carolyn P on May 24, 2017 3:00:00 AM

It’s Friday night and you could be out with your friends right now, but instead you’re tucked away in a dark little room filled with microscopes. Spending the evening in the lab seemed like a good choice at the time because you were certain this immunohistochemistry was going to reveal some small - but important - mystery of the universe to you. But now you’re sitting here, cursing the universe and everyone in it, because all you see when you stare down into the scope is some indistinct fuzziness. And did the controls work - meh - who’s to say? There’s no sugar coating it. It’s a fail.

READ MORE >

Topics: Protocols, IHC, Companion Reagents, techniques

Successful Immunofluorescence: Antibody Dilution and Incubation Conditions


Posted by Ken B on May 10, 2017 3:01:00 AM

Part four of a series on immunofluorescence techniques. Check out previous posts on Validation, Experimental Controls, and Fixation/Permabilization

After your samples have been prepared, it's time to incubate them with a well-validated antibody,  the workhorse of immunofluorescence. If you are a seasoned pro at IF experiments, you are probably used to checking the antibody datasheet (or web page) for the recommended dilution. But have you ever wondered where those recommendations come from?
READ MORE >

Topics: Protocols, Antibody Performance, Primary Antibodies, IF-IC, techniques

Successful Immunofluorescence: Fixation and Permeabilization


Posted by Ken B on Mar 15, 2017 3:00:00 AM

Part Three of a four-part series on Immunofluorescence. Check out The Importance of Validation and Experimental Controls.

The performance of an antibody is a crucial determinant in getting reliable immunofluorescence (IF) results. Equally important is the preparation of the biological sample - cells or tissue used in your experiments - before any antibodies are introduced. The fixation and permeabilization of your samples are key steps that can determine your experiment’s failure or success. The ideal fixative preserves a “life-like” snapshot while quickly stopping the degradative process of autolysis by crosslinking and inhibiting endogenous enzymes. This post provides examples of how different antibodies perform at their best using different protocols.

READ MORE >

Topics: Protocols, Antibody Performance, IF-IC, techniques, Fixation

Successful Immunofluorescence: Experimental Controls


Posted by Ken B on Mar 8, 2017 3:00:00 AM

 Part two of a four-part series on Immunofluorescence. Check out our posts on Validation and Fixation and Permeabilization

After months of hard work, your research has honed in on a hypothesis you can test with immunofluorescence (IF). You've chosen antibodies and performed pilot IF experiments (see The Importance of Validation), and the localization of the protein appears reasonable. But how can you be sure the IF data you've acquired represents real biological phenomena? We present two examples of experimental controls in this post.

READ MORE >

Topics: Protocols, Antibody Performance, Primary Antibodies, Antibody Validation, IF-IC, Post Translational Modification, Reproducibility

Successful Immunofluorescence: The Importance of Validation


Posted by Ken B on Mar 1, 2017 3:00:00 AM

Part one of a four-part series on Immunofluorescence. Check out Experimental Controls and Fixation and Permeabilzation.

After months of hard work, your research has zeroed in on a hypothesis you can test with immunofluorescence (IF). But now you have to make a choice. How do you decide which antibody to use to get reliable IF results? How do you know if the images are accurately reporting the target's localization? We explore some considerations in this post.

READ MORE >

Topics: Protocols, Antibody Performance, Primary Antibodies, Antibody Validation, IF-IC, Reproducility

3 Questions to ask before starting your next immunofluorescence experiment.


Posted by Liana G on Oct 19, 2016 3:00:00 AM

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

  • Is your antibody specific?
  • Is your antibody supported by an optimized IF protocol?
  • Is your antibody performing consistently?

Let's take them each in turn...

READ MORE >

Topics: Protocols, Primary Antibodies, Antibody Validation, IF-IC, Cell Biology

3 Questions to ask before starting your next immunofluorescence experiment.


Posted by Carolyn P on Sep 7, 2016 3:00:00 AM

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

  • Is your antibody specific?
  • Is your antibody supported by an optimized IF protocol?
  • Is your antibody performing consistently?

Let's take them each in turn...

READ MORE >

Topics: Protocols, Primary Antibodies, Antibody Validation, IF-IC, Cell Biology, techniques

Webinar - Multiplex Immunohistochemistry (IHC)


Posted by Liana G on Jun 8, 2016 3:00:00 AM

Analysis of Immune Checkpoint Control Protein Co-expression in Breast and Ovarian Cancer Using Novel Rabbit Monoclonal Antibodies and Multiplex IHC

With an increasing number of biomarkers and, often, limited availability of biopsy material, there is a growing need for multiplexed assays for both research and clinical purposes. IHC based solutions are particularly attractive in the field of immuno-oncology, as maintaining spatial context within the tumor microenvironment provides meaningful and potentially actionable information.

Watch the video below.

 

READ MORE >

Topics: Protocols, IHC, Cancer Immunology, Cancer Research, mIHC

Webinar - Multiplex Immunohistochemistry (IHC)


Posted by Liana G on Jan 13, 2016 3:00:00 AM

Analysis of Immune Checkpoint Control Protein Co-expression in Breast and Ovarian Cancer Using Novel Rabbit Monoclonal Antibodies and Multiplex IHC

With an increasing number of biomarkers and, often, limited availability of biopsy material, there is a growing need for multiplexed assays for both research and clinical purposes. IHC based solutions are particularly attractive in the field of immuno-oncology, as maintaining spatial context within the tumor microenvironment provides meaningful and potentially actionable information.

Watch the video below.

 

READ MORE >

Topics: Protocols, IHC, Cancer Immunology, Cancer Research, mIHC

Subscribe to Email Updates

Recent Posts