Sodium Citrate Antigen Retrieval Buffer (50X)

Sample type

FFPE tissues

Assay type

Immunohistochemistry

Concentration

50x

pH (1x)

6.0

Publications

11

Cat no : PR30001

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Validation Data Gallery

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  IHC analysis of rat brain tissue with Proteintech’s TDP-43 rabbit recombinant antibody (80001-1-RR). Heat-induced epitope retrieval was performed using Sodium Citrate Antigen Retrieval Buffer (PR30001).

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  IHC analysis of rat kidney tissue with Proteintech’s AQP2 rabbit polyclonal antibody (29386-1-Ap). Heat-induced epitope retrieval was performed using Sodium Citrate Antigen Retrieval Buffer (PR30001).

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Product Information

PR30001 is a citrate 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 PR30001 (Sodium Citrate Antigen Retrieval Buffer 50x) to prepare a 1x solution by adding the concentrated buffer to ddH2O. For example, to make 500 mL of 1x working solution, dilute 10 mL PR30001 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 1x 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.

Sodium Citrate 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

PR30001, Sodium Citrate Antigen Retrieval Buffer (50X), Proteintech, IL, USA

Publications

Application	Title
IHC	Drug Resist Updat; Lansoprazole (LPZ) reverses multidrug resistance (MDR) in cancer through impeding ATP-binding cassette (ABC) transporter-mediated chemotherapeutic drug efflux and lysosomal sequestration; Authors - Ning Ji
IF	J Transl Med; MORN2 regulates the morphology and energy metabolism of mitochondria and is required for male fertility in mice; Authors - Yining Liu
	Cells; Deficiency of the Tmem232 Gene Causes Male Infertility with Morphological Abnormalities of the Sperm Flagellum in Mice; Authors - Xiuqing He
IHC	Int J Mol Sci; Cyp19a1a Promotes Ovarian Maturation through Regulating E2 Synthesis with Estrogen Receptor 2a in Pampus argenteus (Euphrasen, 1788); Authors - Chunyang Guo
	Front Mol Biosci; Identification of the m6A RNA Methylation Regulators WTAP as a Novel Prognostic Biomarker and Genomic Alterations in Cutaneous Melanoma.; Authors - Zi-Yi Feng
	Cancer Biol Ther; VAMP8 suppresses the metastasis via DDX5/β-catenin signal pathway in osteosarcoma; Authors - Shuo Yang