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The Truth About Antibodies used for Research

The Problem with Antibodies used for Research

Antibodies used for Research in academic, industrial and therapeutic fields are universal reagents in molecular biology assays. Without them, the entire field of medical research could collapse. However, there are many crappy antibodies out there – this is well known. Moreover, scientists often disregard the need to investigate and validate the antibodies they’ve purchased.

”Scientists often know, anecdotally, that some antibodies in their field are problematic, but it has been difficult to gauge the size of the problem across biology as a whole. Perhaps the largest assessment comes from work published by the Human Protein Atlas, a Swedish consortium that aims to generate antibodies for every protein in the human genome. It has looked at some 20,000 commercial antibodies so far and found that less than 50% can be used effectively to look at protein distribution in preserved slices of tissue.”

Antibodies used for Research : The Reproductibility Crisis

In 2015, Monya Baker wrote about the Reproductibility Crisis with Antibodies used for Research in Nature News in 2015. She describes that the results of many biomedical experiments cannot be reproduced and that the conclusions based on them may be unfounded. Notably, it appears that a lot of scientists shop on price and speed of delivery, which are variables that don’t always correlate well with reagent quality.

In our opinion, trusting what’s on label may or may not be the best strategy for any scientist. With the new era of scientists relying on web-based research hoping to find, what they see as reliable sources, the best and cheapest antibody within 10 min of Google searches, the problem with Antibodies used for Research is not about to be solved. Yo a lesser extent, there’s an analogy to be made with primer design tools for DNA amplification by PCR. Nowadays, young scientists rarely design primers by hand and rely on algoryth-based design softwares. They’re often good of course, but they may not entirely consider what the scientist wishes to probe by PCR using their available tools and the nature of the template DNA.

Private labelling, OEM and dilution : Who says the Truth about Antibodies used for Research?

Now comes the difficult part. The Truth about Antibodies used for Research is that you never know where your antibody really is manufactured. The problem is that the research antibody market has grown so much in the last 3 decades that a reputation for quality is becoming part of some suppliers’ business plans. But most notably, many want to be able to offer the largest portfolio or Antibodies used for Research as possible.


What is Private Labelling?

Private-label products or services, also known as “phantom brands”, are typically those manufactured or provided by one company for offer under another company’s brand. Private-label goods and services are available in a wide range of industries from food to cosmetics to web hosting and of course, biomedical reagents such as Antibodies. They are often positioned as lower-cost alternatives to regional, national or international brands (think about the low-cost Biotech companies), although recently some private label brands have been positioned as “premium” brands to compete with existing “name” brands. The opposite may also be true, i.e re-labelling an inferior product into a ”premium” new product.


OEM and Antibody Dilution

OEM stands for Original Equipment Manufacturer Hence, an OEM is a company that makes a part or subsystem that is used in another company’s end product. For instance, if Acme Manufacturing Co. makes power cords that are used on IBM computers, Therfore, Acme is an OEM. By similarity, if company X manufactures the antibodies used in company Y’s ELISA kits, X is the OEM supplier of company Y.

Notably, The resale of diluted antibodies is probably the most horrifying practice, but it exists. You can see this from big name companies in Example 1 and Example 3 just below.


Are these Practices Wrong?

Don’t get me wrong here. Private labelling and OEM are normal practices in all spheres of business. They are actually good practice and encourage competition. The problem with the field of Antibodies used for Research is that most companies (especially the big ones) wish to offer scientist with the largest portfolio possible. Their strategy is thus aimed at making scientists buy every antibody from them. Basically, they want to become richer of your back. The real experts are actully the OEM/private label companies behind these antibodies. Most of the time, if you can find them, you can purchase the orginal antibody from these companies and save tens or hundreds of dollars.

Those companies who ”pre”-dilute antibodies indeed have questionnable practices…


But it’s not the end of the Problem!

Continue reading…

The Mixed Portfolio of Companies – Antibodies used for Research

Consider the following:

  • Company B supplies antibodies to Company A
  • Company D supplies antibodies to Company C
  • Company D supplies antibodies to Company A
  • Company C supplies antibodies to Company A

Portfolio of Antibodies used for Research of Company A = Portolio A + B + (C+D) + D

This may not always be trivial for all large suppliers, but it happens and confuses research scientists (and even product distributors such as Civic Bioscience). Often, orginal images are replaced with customer data or in-house data, in return causing even more confusion.

Companies and Antibodies used for Research

Yes! Some Companies are Honest

The Original Antibody Manufacturer are, most often, honest and real scientists.

The History behind EnCor Biotechnology

EnCor Biotechnology Inc. was founded by Dr. Gerry Shaw, who was born in Nottingham, England sometime late in the last millennium.

Prior to working in EnCor Biotechnology, employees were full-time researchers working in neuroscience, structural biology and cell biology. We found that good antibody reagents were essential to get any kind of quality data and we also found that many commercial reagents were not of high enough quality to be useful. In fact, many of them were a waste of both time and money. As a result, we often made our own reagents which then became popular with other researchers.

At EnCor we are building on this experience by generating further reagents which we extensively characterize. We are confident that these reagents will work as advertized in the assays we have described. In most cases the antibodies are selected from a panel of reagents. Thus they are not simply the result of one attempt to make an antibody to a particular protein.

In consequence, many of our newer antibodies became popular with collaborators and several found their way to commercial vendors, initially Chemicon (now owned by EMD Millipore), then Novus Biologicals, Novocastra (now owned by Leica), Sigma-Aldrich and Zymed (now owned by Life Technologies).

Importantly, EnCor Biotechnology uses Research Resource Identifiers (RRID numbers; i.e AB_2572391) to identify their antibodies. With RRIDs you can track the original manufacturer of the product itself!

EnCor Biotechnology Logo

New Supplier!

Civic Bioscience is proud to announce and welcomes EnCor Biotechnology as a new supplier of high-quality Primary Antibodies, ELISA kits and Purified Proteins!

What can we do about the Crisis with Antibodies used for Research?

VALIDATE OUR ANTIBODIES

Validate regardless of what the manufacturer claims.

It will benefit all the scientific community!


EnCor Biotechnology uses Research Resource Identifiers (RRID numbers; i.e AB_2572391) to identify their antibodies. Such a practice from other vendors is strongly encouraged.

NOW ENJOY THE SHOW! (Examples below)

Here are 4 examples of Antibodies available on the market, each from 4 different suppliers and manufactured by…

  1. Fox3/NeuN –> RRID # AB_2572267
  2. GFAP –> RRID # AB_2572310
  3. MAP2 –> RRID # AB_2138173
  4. Ubiquitin –> RRID # AB_2572391

Example 1 : Fox3/NeuN Monoclonal Antibody (clone 1B7)

Where Does it Really Come From?
RRID # AB_2572267

Abcam

Anti-NeuN antibody 1B7 Neuronal Marker (ab104224)
425$ + 25$ S&H /100 ul
  • Immunogen: Recombinant fragment corresponding to Human NeuN aa 1-100 (N terminal) expressed in and purified from E. coli.
  • Isotype: IgG2a
  • Reactivity: Mouse, Rat, Goat, Chicken, Cow, Cat, Dog, Human, Pig, Sea urchin, Common marmose
  • Ab Concentration: Affinity purified at 1 mg/mL
  • Applications: IHC-FoFr, IHC-P, WB, ICC/IF
  • Suggested Dilutions: WB: 1:1,000-1:2,000. ICC/IF and IHC: 1:1,000.
  • Abcam rating: 5 stars (10 reviews)
  • Fox3/NeuN Antibody - MCA-1B7 Image 2

Abgent

FOX3 (NeuN) Mouse Monoclonal Antibody 1B7
(Froggabio) 704$ + S&H /100 ul
  • Immunogen: Not specified
  • Isotype: IgG2a
  • Reactivity: Bovine, Human, Mouse, Rat
  • Ab Concentration: not specified
  • Applications: WB, IF
  • Suggest Dilutions: Western blot: 1:2,000. IF 1:500 (diluted antibody?)

BioLegend

Purified anti-FOX3 (NeuN) Antibody (#834501; 1B7)
327.24$ + S&H/100 ul
  • Immunogen: Not specified
  • Isotype: IgG2a
  • Reactivity: Human, Cow, Pig, Mouse, Rat, and other mammals
  • Ab Concentration: 1 mg/mL of ???
  • Applications: WB, ICC/IF, IHC
  • Suggest Dilutions: Western blot: 1:500-1,000 (diluted antibody?). ICC/IF and IHC: 1:1,000.
  • WB image: not shown

Example 2 : GFAP Rabbit Polyclonal Antibody

Who Really Makes It?
RRID # AB_2572310

Abcam

Anti-GFAP antibody (ab7260)
441$ + 25$ S&H /50 ul
  • Immunogen: Full length native protein (purified) corresponding to Human GFAP.
  • Isotype: IgG
  • Reactivity: Mouse, Rat, Cat, Human, Common marmoset
  • Ab Concentration: 100 µg at 1 mg/ml
  • Applications: IHC-FoFr, IHC-Fr, ICC/IF, WB, IHC-P, ICC
  • Suggested Dlutions: WB: 1/10 000; ICC/IF: 1/1000-1/5000
  • Abcam rating: 5 stars (75 reviews)
  • Glial Fibrillary Acidic Protein

Novus

GFAP Antibody, NB300-141
369$ + 29$ S&H /50 ul
  • Immunogen: GFAP purified from bovine spinal cord .(not true! this is for EnCor GFAP, chicken polyclonal, Cat# CPCA-GFAP)
  • Isotype: Rabbit IgG
  • Reactivity: Human, Mouse, Rat, Porcine, Bovine, Canine, Chicken, Feline, Guinea Pig, Rabbit
  • Ab Concentration: not specified
  • Applications: WB, ICC/IF, IHC
  • Suggested Dilutions: WB: 1:5,000; IF/ICC and IHC 1:1,000-1,5,000
  • Glial Fibrillary Acidic Protein

ThermoFisher

GFAP Polyclonal Antibody, # PA3-16727
544$ + S&H /50 ul
  • Immunogen: Native GFAP purified from bovine Spinal Cord.(not true! this is for EnCor GFAP, chicken polyclonal, Cat# CPCA-GFAP)
  • Isotype: Rabbit IgG
  • Reactivity: Bovine, Dog, Chicken, Cat, Guinea pig, Human, Mouse, Pig, Rabbit, Rat
  • Ab Concentration: not specified
  • Applications: WB, ICC/IF, IHC
  • Suggest Dilutions: WB: 1:5 000. ICC/IF and IHC: 1:1,000-1:5000
  • Glial Fibrillary Acidic Protein

Example 3 : Microtubule-Associated Protein 2 (MAP2) Chicken Polyclonal Antibody

Guess Who Makes It?
RRID # AB_2138173

Abcam

Anti-MAP2 antibody (ab5392)
439$ + 25$ S&H/25 ul
  • Immunogen: Full length native protein (purified) corresponding to Cow MAP2 (not accurate)
  • Isotype: IgY
  • Reactivity: Human, horse, cow, pig, chicken, rat, mouse
  • Ab Concentration: Concentrated IgY preparation
  • Applications: ELISA, IHC-Fr, IHC-FoFr, IHC-P, WB, ICC/IF, IHC (PFA fixed)
  • Suggested Dilutions: WB: 1:20,000-100 000 IF/ICC and IHC: 1:10,000-1:20,000.
  • Abcam rating: 5 stars (31 reviews)

Novus

MAP2 Antibody; NB300-213
307$ + 29$ S&H /50 ul
  • Immunogen: Purified MAP2 isolated from bovine brain (not accurate)
  • Isotype: chicken polyclonal IgY
  • Reactivity: Human, Mouse, Rat, Porcine, Bovine, Canine, Chicken, Feline, Guinea Pig, Rabbit
  • Ab Concentration: Ammonium sulfate precipitation
  • Applications: WB, ICC/IF, IHC, IHC-Fr
  • Suggested Dilutions: WB: 1:10000-1:20000; IF/ICC: 1:5000-1:10000 and IHC 1:1,000-1,5,000 (DILUTED ANTIBODY!)
  • Microtubule Associated Protein 2 image 1

EMD Millipore

Anti-MAP2 Antibody, AB5543
403$ + S&H/50 ul
  • Immunogen: Purified MAP2 isolated from bovine brain (not accurate)
  • Isotype: chicken polyclonal IgY
  • Reactivity: Rat
  • Ab Concentration: Ammonium sulfate precipitation
  • Applications: WB, ICC/IF, IHC.
  • Suggested Dilutions: WB: 1:10000-1:20000; IF/ICC: 1:5000-1:10000 and IHC not specified (DILUTED ANTIBODY!)

Example 4 : Ubiquitin Ub-1 Mouse Monoclonal Antibody

Guess Who?
RRID # AB_2572391

Abcam

Anti-Ubiquitin antibody (Ubi-1) (ab7254)
439$ + 25$ S&H /50 ul
  • Immunogen: Purified ubiquitin conjugated with glutaraldehyde to KLH
  • Isotype: Mouse IgG1
  • Reactivity: Mouse, Rat, Chicken, Cow, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Drosophila C Virus
  • Ab Concentration: Supplied as an aliquot of 1 mg/mL of affinity purified antibody.
  • Applications: IHC-P, WB, ELISA, ICC/IF, IHC-Fr
  • Suggested Dilutions: WB: 1:5 000. IF/ICC, IHC: 1:500-1:1 000
  • Western blot - Anti-Ubiquitin [Ubi-1] antibody (ab7254)
  •  

Invitrogen

Ubiquitin Monoclonal Antibody (Ubi-1), MA1-10035
544$ + S&H /100 ul
  • Immunogen: Purified ubiquitin conjugated with glutaraldehyde to KLH
  • Isotype: Mouse IgG1
  • Reactivity: Bovine, C. elegans, Chicken, Fruit fly, Horse, Human, Mouse, Pig, Rat
  • Ab Concentration: Not mentionned… of ascites (typo)
  • Applications: WBICC/IF, IHC-Fr,IHC-P
  • Suggested Dilutions: WB: 1:5 000. IF/ICC, IHC: 1:5000 (clearly a typo)
  • Ubiquitin Antibody - MCA-Ubi-1 Image 2

EMD Millipore

Anti-Ubiquitin Antibody, clone Ubi-1 (aka 042691GS)
392$ + 25$ S&H /100 ul
  • Immunogen: Purified ubiquitin (bovine RBC) covalently coupled to KLH using glutaraldehyde.
  • Isotype: Mouse IgG1
  • Reactivity: H, M, R
  • Ab Concentration: Supplied as an aliquot of ascitic fluid.
  • Applications: IHC(P), IP, WB
  • Suggested Dilutions: WB: 1:250-1000. IF/ICC, IHC: 1:250-1:1,000 (ascites, not purified OK)
  • No WB image

3 TRUE Antibody Manufacturers distributed by Civic Bioscience in Canada

The remaining suppliers are mixed manufacturers/private label (Elabscience and Enogene)


Part of our mission is to offer scientists products from the original manufacturers as much as possible. In return, this offers better pricing, quality as well as better science. We believe in staying TRUE to science.

 

Choose your Antibodies Wisely

ENTER Promo Code TRUTH2017 to receive a 20$ discount on any Primary Antibody 🙂

One use per person. Valid until August 30th, 2017