Specific protein block

Manufactured by Leica

Sourced in United Kingdom

The Specific Protein Block is a laboratory product designed to block non-specific protein binding in immunohistochemical and Western blot applications. It serves to prevent the occurrence of background signals, thus improving the specificity and sensitivity of the target protein detection.

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Lab products found in correlation

Zen 2, by Zeiss (2 mentions) Harris hematoxylin, by Leica (2 mentions) Axiocam 503 color, by Zeiss (1 mentions) Epitope retrieval solution ph 9, by Agilent Technologies (1 mentions) Cd4 4sm95, by Thermo Fisher Scientific (1 mentions) Axiocam 503 color camera, by Zeiss (1 mentions) Axio scope a1, by Zeiss (1 mentions) Epitope retrieval solutions ph6, by Agilent Technologies (1 mentions)

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Protein block (14 citations)

3 protocols using specific protein block

Immunohistochemical Analysis of Tumor-Infiltrating T Cells

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The tumors were fixed in 10% neutral buffered formalin and included in paraffin. 4μm-thick tissue sections obtained from paraffin embedded tissues were deparaffinized, rehydrated and stained with H&E to define tumor histotypes. Immunohistochemistry was performed using a polymer detection method. Briefly, the antigen retrieval was performed using Novocastra Epitope Retrieval Solution pH 9 in a PT Link Dako pre-treatment module at 98°C for 30 minutes. Subsequently, the sections were brought to room temperature and washed in PBS. After neutralization of the endogenous peroxidase with 3% H2O2 and Fc blocking by a specific protein block (Novocastra UK), the samples were incubated with primary antibodies. For rat anti-mouse monoclonal CD8a (4SM15) or CD4 (4SM95) 1:100 pH9 (eBioscience) antibodies, the staining was revealed using goat anti-rat IgG (H&L) specific secondary antibody 1:500 (Novex by Life Technologies) and AEC (3-Amino-9-ethylcarbazole) substrate-chromogen. Slides were analyzed under a Zeiss Axioscope A1 and microphotographs were collected using a Zeiss Axiocam 503 Color with the Zen 2.0 Software (Zeiss). Quantitative IHC data for CD8 and CD4 marker was calculated by counting the number of CD8+ or CD4+ cells in five fields at 40X magnification.

Simula L., Pacella I., Colamatteo A., Procaccini C., Cancila V., Bordi M., Tregnago C., Corrado M., Pigazzi M., Barnaba V., Tripodo C., Matarese G., Piconese S, & Campello S. (2018). Drp1 Controls Effective T Cell Immune-Surveillance by Regulating T Cell Migration, Proliferation, and cMyc-Dependent Metabolic Reprogramming. Cell Reports, 25(11), 3059-3073.e10.

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Immunohistochemical Staining Protocol for Tissue Analysis

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Immunohistochemistry (IHC) was performed using a polymer detection method. Briefly, tissue samples were fixed in 10% buffered formalin and paraffin-embedded. The, the 4-µm thick tissue sections were deparaffinized and rehydrated.
The antigen unmasking technique was performed using Novocastra Epitope Retrieval Solutions pH = 9 or pH = 6 in a PT Link Dako pre-treatment module at 98 °C for 30 min. The sections were then brought to room temperature (RT) and washed in PBS. After neutralization of the endogenous peroxidase with 3% H₂O₂ and Fc blocking by a specific protein block (Novocastra, Newcastle, UK), the samples were incubated for 1h with the primary antibodies at RT. Primary antibodies were listed in Table 1.
Staining was revealed by polymer detection kit (Novocastra) and 3-Amino-9-ethylcarbazole (AEC) Dako substrate chromogen. The slides were counterstained with Harris hematoxylin (Novocastra). Slides were analyzed under a Zeiss Axioscope A1, and microphotographs were collected using a Zeiss Axiocam 503 Color camera with the Zen 2.0 Software (Zeiss, Oberkochen, Germany).
IHC was evaluated based on the intensity of staining and scored as grade 0 (negative), grade 1 (weak), grade 2 (moderate) or grade 3 (strong).

Belmonte B., Cancila V., Gulino A., Navari M., Arancio W., Macor P., Balduit A., Capolla S., Morello G., Vacca D., Ferrara I., Bertolazzi G., Balistreri C.R., Amico P., Ferrante F., Maiorana A., Salviato T., Piccaluga P.P, & Mangogna A. (2021). Constitutive PSGL-1 Correlates with CD30 and TCR Pathways and Represents a Potential Target for Immunotherapy in Anaplastic Large T-Cell Lymphoma. Cancers, 13(12), 2958.

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Immunohistochemical Tissue Analysis Protocol

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Tissue samples were fixed in 10% buffered formalin and paraffin embedded. Four-micrometersthick tissue sections were deparaffinized and rehydrated. The antigen unmasking technique was performed using Novocastra Epitope Retrieval Solutions pH6, pH 9, and pH 8 in a PT Link Dako pre-treatment module at 98°C for 10 minutes. The sections were then brought to room temperature and washed in PBS. After neutralization of the endogenous peroxidase with 3% H 2 O 2 and Fcblocking by a specific protein block (Novocastra UK), the samples were incubated overnight at 4°C with the primary antibodies. IgG from normal rabbit sera were used as negative control. Staining was revealed by the Horseradish Peroxidase (HRP) polymer detection kit (Novocastra, Code RE7280-K). The sections were counterstained with Harris hematoxylin (Novocastra) and analysed under a Leica DMD108 optical digital microscope (Leica Microsystems, Germany).

Agostinis C., Rami D., Zacchi P., Bossi F., Stampalija T., Mangogna A., Amadio L., Vidergar R., Vecchi Brumatti L., Ricci G., Celeghini C., Radillo O., Sargent I, & Bulla R. (2018). Pre-eclampsia affects procalcitonin production in placental tissue. American journal of reproductive immunology (New York, N.Y. : 1989), 79(4).

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