Antigen Retrieval Method
BioGenex is the inventor of Antigen Retrieval enabling technology. Antigen Retrieval is an effective way of unmasking antigenic epitopes on the surface of formalin-fixed, parraffin-embedded tissue sections using microwave heating. Covered by exclusive patents issued to BioGenex, this method to recover antigenicity in formalin-fixed, paraffin-embedded tissue sections has been routinely practiced in laboratories throughout the world.
The Antigen Retrieval technique breaks the formalin induced cross-linking bonds between epitopes and unrelated proteins, thereby allowing better penetration of antibody and accessibility of epitopes.
Advantages of the Antigen Retrieval Technique
- Enhanced exposure of antigenic epitopes on the surface of the tissue section
- Reduces time for primary antibodies incubation
- Consistent and reliable staining quality
- Eliminates false negative staining results in FFPE tissue sections
- Ease-of-use
Different antibodies require different conditions for Antigen Retrieval.
BioGenex offers several types of Antigen Retrieval solutions:
1. Antigen-Retrieval (AR) Solutions – For Manual Use
| Product |
Method |
Features & Recommended Use |
| Citra |
Microwave, 95-100°C |
PH~6, excellent for most BioGenex Antibodies* |
| Citra Plus |
Microwave, 95-100°C |
Enhanced formulation PH~6, for antibodies such as Estrogen Receptor (clone ER88), HSP27 (G3.1) and CDX-2 (CDX2-88)* |
| AR-10 |
Microwave, 95-100°C |
Tris-Based, high PH-10, for antibodies such as Caldesmon (clone h-CD), CD3 (PS1), c-myc (9E10) & GLEPP1 (5C11)* |
| H&E |
Microwave or Room Temp. 25-100°C |
Best for burnt, overfixed or dried FFPE tissues, over-DeCal (bone marrow biopsies) and fragile/over processed specimens (e.g. needle biopsies). Can be used at room temp. for some frozen tissue sections and tissues with freezing artifacts. |
| DeCal |
Room Temp. 20-25°C |
For acid-decalcified bone marrow & formalin-fixed tissues embedded in paraffin or celloidin. |
* See datasheets for BioGenex recommended Antigen Retrieval for each specific antibody.
2. Easy & Enhanced Antigen-Retrieval (EZ-AR) Solutions – For Manual Use
Features & Benefits
- Unique superheating properties increase the availability of antigenic epitopes in tissues.
- Short and standardized protocols for all BioGenex antibodies eliminates guesswork in optimizing protocols
- Fast uniform heating and cooling of solutions reduces tissue pretreatment time
- Non-hazardous, non-flammable and odorless makes them safe and exo-friendly
| Product |
Method |
Features & Recommended Use |
| EZ-AR 1 |
EZ-Retriever or Microwave, 107°C |
Citric acid based, PH~6, excellent for most BioGenex Antibodies*. |
| EZ-AR 2 |
EZ-Retriever or Microwave, 107°C |
EDTA based, PH~8.5, for antibodies such as Ki67 (EP5), CD5 (EP2952) and NGF Receptor (EP1039Y)*. |
| EZ-AR 3 |
EZ-Retriever or Microwave, 95-100°C |
Citric acid based, PH~6, for antibodies such as Estrogen Receptor (clone ER88), HSP27 (G3.1) and CDX-2 (CDX2-88)* |
| EZ-AR 4 |
EZ-Retriever or Microwave, 95-100°C |
Tris based, PH~10, for antibodies such as Caldesmon (clone h-CD), CD3 (PS1), c-myc (9E10) & GLEPP1 (5C11)* |
* See datasheets for BioGenex recommended Antigen Retrieval for each specific antibody.
3. Xmatrx Antigen-Retrieval (EZ-AR Elegance) Solutions – For use with Xmatrx Automation
Features & Benefits
- Optimized for Xmatrx with standardized protocols for all BioGenex antibodies.
- Reaches 107°C without boiling to minimize evaporation and preserve morphology.
- Non-hazardous, non-flammable and odorless makes them safe and eco-friendly
| Product |
Method |
Features & Recommended Use |
| EZ-AR 1 Elegance |
Xmatrx Automation Works at 100-105°C |
Citric acid based, PH~6, excellent for most BioGenex Antibodies*. |
| EZ-AR 2 Elegance |
Xmatrx Automation Works at 100-105°C |
EDTA based, PH~8.5, for antibodies such as Ki67 (EP5), P27 (Y236) and P53 Protein (DO7)*. |
| EZ-AR 3 Elegance |
Xmatrx Automation Works at 95-100°C |
Citric acid based, PH~6, for antibodies such as Estrogen Receptor (clone ER88), HSP27 (G3.1) and CDX-2 (CDX2-88)* |
| EZ-AR 4 Elegance |
Xmatrx Automation Works at 95-100°C |
Tris based, PH~10, for antibodies such as Caldesmon (clone h-CD), CD3 (PS1), c-myc (9E10) & GLEPP1 (5C11)* |
* See Xmatrx protocols for BioGenex recommended Antigen Retrieval for each specific antibody
Breast Carcinoma tissue stained with Anti-Progesterone Receptor [PR88] MAb, using AEC chromogen without Antigen Retrieval.
Breast Carcinoma tissue stained with Anti-Progesterone Receptor [PR88] MAb, using AEC chromogen with Antigen Retrieval using Citra.
Breast Carcinoma tissue stained with Anti-Estrogen Receptor [ER88] MAb, using DAB chromogen without Antigen Retrieval
Breast Carcinoma tissue stained with Anti-Estrogen Receptor [ER88] MAb, using DAB chromogen with Antigen Retrieval using Citra Plus.
Tonsil tissue stained with anti-CD3 MAb using DAB chromogen without Antigen Retrieval using AR-10.
Tonsil tissue stained with anti-CD3 MAb using DAB chromogen with Antigen Retrieval using AR-10.
Burnt kidney tissue stained after standard pre-treatment.
Nuclear data restored by microwave heating in H&E Solution.
Enzymes for Tissue Digestion
Some tissues require the use of enzymatic pre-treatment before staining to achieve standardized results depending on the antibodies and their different incubation and pre-treatment requirements. Each kit contains three or four vials of lyophilized enzyme powder and 15 ml of reconstitution buffer, enabling you to make fresh enzyme solutions as needed.
- The Trypsin and Pepsin kits contain well-established enzymes suitable for routine pre-treatment at 37°C.
- Protease XXIV kits contain a universal digestive agent that allows for fast and effective pre-treatment at room temperature.
- The diastase (Alpha-amylase) catalyzes breakdown of starch. Diastase is often used prior to periodic acid-Schiff (PAS) staining (See Special Stains section) and is useful as an aid in the evaluation of glycogen storage disease.
- Proteinase K in a ready to use (RTU), RNase-free solution.
Leiomyoma tissue stained with Factor VIII MAb using AEC chromogen following Pepsin Pre-treatment.
Colonic adeno carcinoma stained with Anti-Cytokeratin 20 MAb following Protease Pre-treatment
Squamous Carcinoma tissue stained with Anti-Cytokeratin (High Molecular Weight) MAb using AEC chromogen following Trypsin Pre-treatment.
De-Waxing Solutions
BioGenex deparaffinization solutions are “one-step” products that simultaneously enable the removal of paraffin and allow rehydration of the tissue with a single reagent. In the past, formalin-fixed, paraffin-embedded tissue sections were deparaffinized with highly toxic, noxious chemicals (i.e. xylene, equivalents). BioGenex, a pioneer in the Immunohistochemistry technology, offers xylene-free products that remove the paraffin from mounted tissue slides easily and rapidly.
- EZ-DeWax Solution is for all BioGenex manual methods.
- X-Dewax Solution is optimized for Xmatrx automation.
Features & Benefits:
- Effectively removes paraffin and allow rehydration of the tissue in one step.
- Reduces deparaffinization time from 45 minutes to 10 minutes.
- Eliminates use of toxic solvents (Xylene) and minimizes hazardous waste.
- Ready to use (RTU) or 2x solutions (to be diluted 1:1 with ethanol) are available.
- EZ-AR Common Solution is for microwave facilitated deparaffinization.
Features & Benefits:
- Conveniently perform deparaffinization and Antigen Retrieval in the same slide tank using microwave heating.
- Quick deparaffinization & rehydration in one step (10 minutes @ 70°C default protocol).
- Reduces the use of alcohol in preparing tissue sections for IHC, ISH, H & E, FISH and Special Stains.
- Eliminates use of toxic solvents (Xylene) and minimizes hazardous waste.
- Optimized for use in EZ-Retriever microwave with BioGenex EZ-AR 1-4 solutions.
