Welcome to the Abcam blog

Protocols for IHC and ICC/IF that you can trust

Are you new to immunohistochemistry or immunofluorescence? Have you thought of trying a new immunostaining method but don't know where to start? To help your research take off, we've collected all our IHC and ICC/IF know-how into a comprehensive protocols page covering among other topics:

Browse IHC and ICC/IF protocols»

 

Protocols for your ChIP and epigenetics needs

Don't know where to start with your epigenetic sample preparation? Our epigenetic protocols and troubleshooting tips will support you during your experiments. Our protocols features popular epigenetic applications such as Chromatin Immunoprecipitation (ChIP), RIP, and CLIP and contain details of all the methods, materials and tips you need to successfully complete your experiments.

Browse epigenetics protocols »

 

Western blot protocol video

Watch our western blot protocol video that takes you through sample preparation, electrophoresis, transfer and detection stages.


Our visual step by step guide to western blotting, created in collaboration with our technical team and laboratory, can accompany our procedure for western blotting protocol. This video takes you through:

Sample preparation

The sample lysate is heated in sample buffer to reduce and denature the proteins in the sample.

Electrophoresis

Lysates are loaded into a polyacrylamide gel and an electric field is applied, which causes proteins to migrate through the gel and separate from one another according to size.

Transfer

Proteins are transferred from the gel to a membrane, where the protein of interest can be detected with a specific antibody.

Detection

The membrane is incubated with antibodies, and an enzymatic or fluorescent detection system is used to visualize the specific protein of interest.


Products for western blotting

First edition protocols book now available - limited stock!

There are over 80 pages filled with color illustrations on how to get the best from our products.

The book contains 21 protocol and troubleshooting guides that have been written by our in-house Science Support team and external collaborators who, between them, have years of experience in the lab. It doesn't matter if you are just starting as a lab technician or have worked in a laboratory for years, this book has an abundance of useful reference material.

Protocols include:

  • Fluorescence
  • Western blot
  • Immunohistochemistry
  • ELISA
  • Flow Cytometry
  • Immunoprecipitation
  • ChIP
  • Buffers and stock solutions

The easiest way to get your hands on a copy is by emailing a protocol book request to us.

You can also pick up a printed version at one of our forthcoming events and tradeshows. Future Abcam events over the next few months are Epigenetics and Stem Cells in Copenhagen, Cancer and Metabolism: Pathways to the Future in Edinburgh and Stochastic Events in Stem Cell Differentiation and Reprogramming in Las Vegas.

Alternatively, if you would rather have a PDF for ease of access on your laboratory computer then you can download the book.

Don't forget your can view all our protocols and troubleshooting tips on the Abcam website.

An introduction to protocols

When it comes to research and science, we all rely on the basics and understanding how proteins and molecules work and react with each other. We know that for undergraduates, and researchers exploring new research tools, starting a new unfamiliar application may be scary.

It would be easy if you had Aladdin's lamp, where with a little rub a wish could come true; "I wish I had all the reagents and antibodies needed for my research." POOF! all the necessary reagents and antibodies appear magically in front of you!

However, that is wishful thinking (no pun intended), it's something you might see on a Sunday morning cartoon. In real life the next best thing you could wish for is a group of scientists with lots of experience to help you step by step with your experiment. Well, you're in luck! At Abcam we have experienced scientists who have sat down and written detailed protocols, guides and tips that can help you perform all the regularly used experimental protocols, plus some uncommon ones too!

Visit our Protocols and Troubleshooting page, it contains many application protocols; from basic cell culture, immunostaining (cells, paraffin sections, frozen sections), ELISA (sandwich, direct, indirect), flow cytometry, immunoprecipitation, Western blot and ChIP applications. All these protocols were compiled and written by our Scientific Support team members, who are dedicated in providing you with comprehensive new protocols. We also have some protocols kindly provided to us by loyal customer and collaborators who are experts in their fields.

You may have heard of an exciting technique called ChIP (Chromatin Immunoprecipitation) that can be used to investigate the interaction between proteins and DNA in animal cells, but do you also know that ChIP can also be used for plant samples? What about overcoming cell culture contamination, using PBS or methanol H2O2 solution for blocking endogenous peroxidase activity, milk or BSA for blocking in WB?

How about little bits of information to help you improve your understanding of how reagents work in your experiment, like:

  • Tween 20 in washing buffer will reduce surface tension and increase antibody binding.
  • Triton-X makes holes in cell membrane and allows antibody to bind to intra-membrane epitope.
  • TBS buffer in WB is ideal when looking at phosphorylation sites. Normal PBS contains phosphates which will interfere with results. Also, using milk which contains Casein, is not suitable for use as a blocking solution when detecting phosphorylated proteins.

At Abcam, we have all this information and much, much more. Also, don't miss out on our vast troubleshooting tips and protocol summary diagrams. All protocols are available as PDFs, so you can download the summary diagrams and refer to them on you work bench as you perform your experiments. Sure beats the college note pads or A4 print outs with lines and lines of boring protocols.

Although we do not have magical lamps to hand out, we sure do have protocols, troubleshooting tips, diagrams, and other information that we know will be useful for you. Not to mention scientific support members around the globe with expertise in almost all research areas to support you. Feel free to contact us anytime, we are waiting to hear from you!

ChIP protocols and new ChIP primers from Abcam

Chromatin Immunoprecipitation (ChIP) enables the mapping of target proteins/histone modifications to specific loci in the genome. Antibodies and protein A beads, which have a higher affinity for some immunoglobulin isotypes, are used to immunoprecipitate chromatin/DNA complexes. The DNA region of interest is determined using quantitative PCR.

Here are six protocols to help you get the best out of your experiment.

ChIP protocols

If you're working in a different application, we now have over 70 protocols, including guides for ELISA, FACS, IP and WB, and Abcam's Scientific support team are adding more all the time!

New ChIP primers - fast, easy, and consistent

We can now provide validated ChIP primers which you can use to:

  • Test antibodies for use in ChIP
  • Control for antibodies already validated in ChIP