Antigen retrieval

Manufactured by Vector Laboratories

Sourced in United States

Antigen retrieval is a laboratory technique used to enhance the detection of target antigens in tissue sections. It is a critical step in immunohistochemistry and other immunodetection methods. The process involves the use of various treatments to unmask the epitopes of target proteins, making them more accessible for antibody binding and subsequent visualization.

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Lab products found in correlation

3 3 diaminobenzidine (dab), by Merck Group (1 mentions) Permanent mounting media, by Vector Laboratories (1 mentions) Horse serum, by Vector Laboratories (1 mentions) Ki67 antibody, by Vector Laboratories (1 mentions) Dm500, by Leica (1 mentions)

Close spelling variants of this product

Antigen retrieval solution (20 citations) Citrate-based antigen retrieval (5 citations) Citrate antigen retrieval buffer (4 citations) Vector Antigen Retrieval Solution (3 citations)

5 protocols using antigen retrieval

Immunohistochemistry of Dicer-Pten DKO Tumors

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Immunohistochemistry (IHC) was performed on archived formalin-fixed, paraffin-embedded Dicer-Pten DKO mouse tumors tissues. Standard xylene deparaffinization, rehydration with a descending series of ethanol solutions, antigen retrieval (Vector Laboratories, Burlingame, CA), and blocking of endogenous peroxidases in 0.3% H₂O₂ were performed as described before [44 (link)]. 3, 3 –diaminobenzidine (DAB) horseradish peroxidase substrate kit was used for color development (Vector Laboratories, Burlingame, CA).

Hua Y., Choi P.W., Trachtenberg A.J., Ng A.C., Kuo W.P., Ng S.K., Dinulescu D.M., Matzuk M.M., Berkowitz R.S, & Ng S.W. (2016). Epithelialization of mouse ovarian tumor cells originating in the fallopian tube stroma. Oncotarget, 7(40), 66077-66086.

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Immunohistochemical Analysis of Pinin in Ovarian Tumors

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Immunohistochemistry (IHC) was performed on a panel of 74 archived formalin-fixed, paraffin-embedded tissues, including 8 normal ovarian tissues and 66 benign, borderline, and malignant ovarian tumors. Standard xylene deparaffinization, rehydration with a descending series of ethanol solutions, antigen retrieval (Vector Laboratories, Burlingame, CA), and blocking of endogenous peroxidases in 0.3% H₂O₂ were performed. The mouse anti-Pinin monoclonal antibody has been described before [22 (link)]. 3, 3 –diaminobenzidine (DAB) horseradish peroxidase substrate kit was used for color development (Vector Laboratories, Burlingame, CA). Staining was graded semiquantitatively by multiplying the proportion of the stained epithelial area (from 0 for absence to 3 for more than 95% of the total epithelial area) with the intensity of the stain (from 0 for negative staining to 3 for strongly positive staining).

Zhang Y., Kwok J.S., Choi P.W., Liu M., Yang J., Singh M., Ng S.K., Welch W.R., Muto M.G., Tsui S.K., Sugrue S.P., Berkowitz R.S, & Ng S.W. (2016). Pinin interacts with C-terminal binding proteins for RNA alternative splicing and epithelial cell identity of human ovarian cancer cells. Oncotarget, 7(10), 11397-11411.

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Immunohistochemical Detection of Protein X

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Briefly, sections were melted, deparaffinized, quenched in methanol-hydrogen peroxide, rehydrated [54 (link), 55 (link)] followed by antigen retrieval (Vector Labs, USA), and blocked with 5% non-fat dry milk (NFDM) containing 5% normal goat serum in PBS (NGS) for 1 hour at room temperature. A 1:100 dilution or 2 μg/ml concentration of IM or PIM antibodies was applied to slides at 4°C overnight. Following three washes, a 1:500 dilution of GαRb HRP was added. After additional washings, brown reaction product was developed using 3, 3′-diaminobenzidine (SIGMA, USA), followed by hematoxylin counterstaining and imaged after mounting.

Pires E.S., D'Souza R.S., Needham M.A., Herr A.K., Jazaeri A.A., Li H., Stoler M.H., Anderson-Knapp K.L., Thomas T., Mandal A., Gougeon A., Flickinger C.J., Bruns D.E., Pollok B.A, & Herr J.C. (2015). Membrane associated cancer-oocyte neoantigen SAS1B/ovastacin is a candidate immunotherapeutic target for uterine tumors. Oncotarget, 6(30), 30194-30211.

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Immunohistochemical Analysis of Ovarian Tumors

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Immunohistochemistry (IHC) was performed on a panel of 21 archived formalin-fixed, paraffin-embedded high-grade malignant tissue samples with known survival data, including 12 high-grade serous, 5 endometrioid, 1 clear cell and 3 mucinous ovarian tumors. Standard xylene deparaffinization, rehydration with a descending series of ethanol solutions, antigen retrieval (Vector Laboratories, Burlingame, CA), and blocking of endogenous peroxidases in 0.3% H₂O₂ were performed. We used both a mouse monoclonal Cyclin A1 antibody B88-2 from BD Biosciences (San Diego, CA, USA) and a rabbit antibody ab118897 from Abcam Inc. (Cambridge, MA). 3, 3–diaminobenzidine (DAB) horseradish peroxidase substrate kit was used for color development (Vector Laboratories, Burlingame, CA). Staining was graded semiquantitatively by multiplying the proportion of the stained epithelial area (from 0 for absence to 3 for more than 95% of the total epithelial area) with the intensity of the stain (from 0 for negative staining to 3 for strongly positive staining).

Huang K.C., Yang J., Ng M.C., Ng S.K., Welch W.R., Muto M.G., Berkowitz R.S, & Ng S.W. (2016). CyclinA1 Expression and Paclitaxel Resistance in Human Ovarian Cancer Cells. European journal of cancer (Oxford, England : 1990), 67, 152-163.

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Immunohistochemical Evaluation of Ki-67

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Collected tissues were processed and sectioned as reported previously [13, 14] . Sections were deparaffinized, rehydrated, and subjected to antigen retrieval (Vector Laboratories) treatment for 20 min followed by blocking the endogenous peroxidase with 3% hydrogen peroxide in methanol for 20 min. Horse serum (Vector Laboratories) was used for blocking the sections for 30 min at room temperature and incubated with Ki-67 antibody (#VP-RM04; Vector Laboratories, 1:100) overnight at 4°C. Sections were washed twice with 1x PBS and treated with biotinylated anti-rabbit/mouse IgG secondary antibody (Vector Laboratories) for 45 minutes, followed by ABC reagent for 30 min. Diaminobenzidine (Vector Laboratories) was used as a substrate to develop the stain. Hematoxylin was used as a counterstain followed by dehydration with alcohol, clearing with xylene, and mounting with permanent mounting media (Vector Laboratories). Stained sections were observed under the microscope (Leica DM500), and images were taken at different magnifications.

Suresh V., Mohanty V., Avula K., Ghosh A., Singh B., Reddy R.R., Suryawanshi A.R., Raghav S.K., Chattopadhyay S., Prasad P., Swain R., Dash R., Parida A., Syed G.H., & Senapati S. (2021). Quantitative proteomics of hamster lung tissues infected with SARS-CoV-2 reveal host-factors having implication in the disease pathogenesis and severity.

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