Tris-EDTA Antigen Retrieval Buffer (50×)

Sample type

FFPE tissues

Assay type

Immunohistochemistry

Concentration

50x

pH (1x)

9.0

Publications

11

Cat no : PR30002

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Validation Data Gallery

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  IHC analysis of mouse pancreas tissue with Proteintech’s Synaptophysin rabbit polyclonal antibody (17785-1-AP). Heat-induced epitope retrieval was performed using Tris-EDTA Antigen Retrieval Buffer (PR30002).

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  IHC analysis of human tonsillitis tissue with Proteintech’s CD8 mouse monoclonal antibody (66868-1-Ig). Heat-induced epitope retrieval was performed using Tris-EDTA Antigen Retrieval Buffer (PR30002).

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Product Information

PR30002 is a Tris-EDTA based solution that can be used for performing heat-induced epitope retrieval (HIER) in immunohistochemistry applications. While treating tissues with formalin and xylene are necessary steps for fixation and deparaffinization in an immunohistochemistry workflow, such treatments often result in protein cross-linking leading to the masking of antigenic sites and the subsequent inhibition of antigen-antibody interactions. Antigen retrieval helps restore the structure of such proteins by breaking the cross-links and unmasking the antigenic sites, thereby making them more accessible to antibodies.

Components

Component	Size
Retrieval Buffer	100 mL

Storage

Stored at 2-8°C. This product is stable for 6 months from the date of receipt.

Usage

1. Prepare slides from tissues sections following routine methods. Deparaffinize tissues with xylene and re-hydrate using a decreasing ethanol gradient.

2. Use concentrated PR30002 (Tris-EDTA Antigen Retrieval Buffer 50x) to prepare a solution by adding the concentrated buffer to ddH2O. For example, to make 500 mL of 1x working solution, dilute 10 mL PR30002 with 490 mL ddH2O.

* 500 mL of prepared 1x antigen retrieval buffer in a 1 L beaker should be suitable for 1 or 2 slide baskets.

3. Heat buffer solution to 95-98°C with a heating element.

* Cover the beaker with a lid or foil to avoid vaporizing/vapor inhalation.

4. Place slide basket into heated antigen retrieval buffer. Maintain this temperature while incubating for 15-20 minutes.

5. Remove the beaker from heat and let it cool to room temperature (takes 35-40 minutes).

6. Perform subsequent steps of quenching, blocking, primary and secondary antibody incubation, signal development, counter staining and mounting before analyzing the slides.

 Tris-EDTA Antigen Retrieval Buffer (50X) User Manual

Application Note

1. Dilute to 1x before use.

2. Calculate how much antigen retrieval buffer will be needed for the experiment to prepare an appropriate amount of 1x working solution. The working solution has a shorter shelf life than the concentrate.

Cited in Article as

PR30002, Tris-EDTA Antigen Retrieval Buffer (50×), Proteintech, IL, USA

Publications

Application	Title
	Cell Mol Biol Lett; The role of NPY2R/NFATc1/DYRK1A regulatory axis in sebaceous glands for sebum synthesis; Authors - Tao Yang
	Front Pharmacol; Deletion of Smooth Muscle Lethal Giant Larvae 1 Promotes Neointimal Hyperplasia in Mice.; Authors - Ya Zhang
	Interact Cardiovasc Thorac Surg; Atrial endocardial expression of von Willebrand factor and thrombomodulin is associated with recurrence after minimally invasive surgical atrial fibrillation ablation.; Authors - Kang An
	Iran J Basic Med Sci; Short-hairpin RNA-mediated suppression of cortactin may inhibit the migration and invasion abilities of endometrial cancer cells by reducing lamellipodia; Authors - Huisi Lin
	Cancer Res; Spatial and Single-Cell Transcriptomics Reveal a Cancer-Associated Fibroblast Subset in HNSCC That Restricts Infiltration and Antitumor Activity of CD8+ T Cells; Authors - Chuwen Li
	Biochem Pharmacol; STING deficiency alleviates ferroptosis through FPN1 stabilization in diabetic kidney disease; Authors - Qin-Xiao Zhao