IHC Enzyme Antigen Retrieval Reagent

Boster’s IHC Enzyme Antigen Retrieval Reagent is a ready to use proteolytic enzyme solution intended to be used as a pretreatment reagent in IHC. Cited in 11 publication(s).

Product Info Summary

SKU AR0022
Pack size 50 mL (for 500-750 assays)
Form SuppliedSolution (liquid)
Applications IHC
PEFF tissues


Physical State Slightly viscous liquid
Pack Size 50 mL (for 500-750 assays)
Form Supplied Solution
Content Multiple proteinases
Composition Proteolytic Enzymes
Volume of Final Solution 50 mL
Enzymatic Activity Protease
Storage Upon receipt store at 4°C. It is stable for one year.
Equivalent N/A
Type Buffers and Mixes
Research Area Molecular Biology, Cell Biology, Histology
Applications IHC
PEFF tissues
Cite This Product IHC Enzyme Antigen Retrieval Reagent (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR0022)

Assay Principle

IHC enzyme antigen retrieval reagent is a ready to use proteolytic enzyme solution intended to be used as a pretreatment reagent in IHC for enzymatic digestion of tissues employed in proteolytic induced epitope retrieval (PIER) techniques applied upon paraffin-embedded formalin-fixed (PEFF) tissue sections on slides following deparaffinization and prior to antibody staining.


The IHC Enzyme Antigen Retrieval Reagent contains multiple proteinases that catalyze proteolysis in tissue samples, breaking the proteins into smaller peptide fractions and amino acids. Enzymatic digestion is commonly used in IHC in the PIER method for antigen retrieval to overcome the loss of immunoreactivity in epitopes caused by the covalent cross-links (methylene bridges) that are formed in tissues during formalin fixation. Proteolysis serves to improve the accessibility of antibodies to target antigens by two main mechanisms: by increasing the penetration of immunoreagents into tissue structures as a result of both tissue digestion and relaxation of the protein structure rigidity, and by restoring the methylene-bridged epitopes. Since proteases exhibit different cleavage site specificity, the combination of multiple proteinases provides a broader spectrum of proteins digestion in a tissue sample. Thus various types of antigenic targets (e.g. protein, glycoprotein, or non-protein, inter- or intracellular antigens) can benefit from this cocktail of digestion enzymes.

Usage and Handling

• The use of enzyme digestion method may destroy some epitopes and tissue morphology. In these cases HIER methods for antigen retrieval should be used.

• When using the enzymatic digestion reagents it is important to not over or under digest. Preliminary experiments should be conducted to determine the optimum incubation time, temperature, and enzyme concentration.

• In general, enzymatic methods are shown to improve IHC staining results less than HIER and are much less reproducible. Their use is primarily recommended for difficult-to-retrieve epitopes when HIER fails to produce satisfactory results.

• IHC Enzyme Antigen Retrieval Reagent is for scientific laboratory research purposes and is not for diagnostic, therapeutic, prophylactic or in vivo applications.

Assay Protocol

1. Add IHC enzyme antigen retrieval reagent directly onto the tissue or cell, and cover the entire tissue or cell completely.

2. Incubate at 37°C for 5-10 minutes.

3. Wash the tissue or cell with PBS.

Product Images

AR0022 has been cited in 11 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Activation of the macroautophagic system in scrapie-infected experimental animals and human genetic prion diseases

Ocular conjunctival inoculation of SARS-CoV-2 can cause mild COVID-19 in rhesus macaques

The pathogenicity of SARS-CoV-2 in hACE2 transgenic mice

Lin D,Xiao Y,Huang B,Wu X,Chen C,Liang Y,Zeng D.O-6-methylguanine DNA methyltransferase is a favorable biomarker with proliferation suppressive potential in Breast Cancer.J Cancer.2020 Sep 1;11(21):6326-6336.doi:10.7150/jca.46466.PMID:33033516;PMCID:PMC75
Species: Human

Tian Y,Gou J,Zhang H,Lu J,Jin Z,Jia S,Bai L.The anti-inflammatory effects of 15-HETE on osteoarthritis during treadmill exercise.Life Sci.2021 Feb 23:119260.doi:10.1016/j.lfs.2021.119260.Epub ahead of print.PMID:33636171.
Species: Rat

Li Z,Huang Z,Zhang H,Lu J,Tian Y,Wei Y,Yang Y,Bai L. P2X7 Receptor Induces Pyroptotic Inflammation and Cartilage Degradation in Osteoarthritis via NF-κB/NLRP3 Crosstalk. Oxid Med Cell Longev. 2021 Jan 16;2021:8868361.doi:10.1155/2021/8868361.PMID:33532039
Species: Rat

Effects of human vascular endothelial growth factor on reparative dentin formation

Activation of the macroautophagic system in scrapie-infected experimental animals and human genetic prion diseases

Song Z, Shah SZA, Yang W, Dong H, Yang L, Zhou X, Zhao D. Front Mol Neurosci. 2017 May 3;10:128. doi: 10.3389/fnmol.2017.00128. eCollection 2017. Downregulation of the Repressor Element 1-Silencing Transcription Factor (REST) Is Associated with Ak...

Bai, Y., Xu, R., Zhang, X., Zhang, X., Hu, X., Li, Y.,..., & Wu, J. (2017). Differential Role of Rapamycin in Epidermis-Induced IL-15-IGF-1 Secretion via Activation of Akt/mTORC2. Cellular Physiology and Biochemistry, 42(5),1755-1768. Advance onli...

Wang, J., Zhang, S., Wu, J., Lu, Z., Yang, J., Wu, H.,…, & Cao, T. (2017). Clinical significance and prognostic value of SOX7 expression in liver and pancreatic carcinoma. Molecular Medicine Reports, 16(1), 499-506. doi: 10.3892/mmr.2017.6660

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