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Endogenous blocking kit

Manufactured by Thermo Fisher Scientific
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The Endogenous Blocking kit is a laboratory product designed to help eliminate non-specific signal in immunohistochemistry (IHC) and other protein detection assays. The kit contains reagents that block endogenous proteins or enzymes that can interfere with the primary antibody-antigen interaction, improving the specificity and accuracy of the results.

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Lab products found in correlation

Immpact dab, by Vector Laboratories (5 mentions) Anti lef1, by Cell Signaling Technology (2 mentions) Anti keratin 10, by BioLegend (2 mentions) Stan dard abc kit, by Vector Laboratories (2 mentions) Mouse anti p53 antibody clone 1c12, by Cell Signaling Technology (2 mentions) Mouse on mouse m o m blocking reagent, by Vector Laboratories (1 mentions)

Close spelling variants of this product

Endogenous Biotin-Blocking Kit

5 protocols using endogenous blocking kit

1

Immunohistochemical Analysis of Key Markers

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For K14, Ki67, K10 and Lef1 immunohistochemistry in human samples, paraffin sections were deparaffinized, rehydrated, followed by antigen unmasking performed for 20 min at 98 °C in citrate buffer (pH 6) using the PT module. Endogenous peroxidase was blocked using 3% H2O2 (Merck) in methanol for 10 min at room temperature. Endogenous avidin and biotin were blocked using the Endogenous Blocking kit (Invitrogen) for 20 min at room temperature. Nonspecific antigen blocking was performed using blocking buffer. Mouse anti-K14 (rabbit, 1/2000, Thermofisher), anti-Ki67 (rabbit, 1/400, Abcam, ab15580), anti-Keratin10 (rabbit, 1/200, Biolegend, ref.90541) and anti-Lef1 (rabbit, 1/100, Cell Signaling, ref.2230) were incubated overnight at 4 °C. Anti-rabbit biotinylated with blocking buffer, Standard ABC kit, and ImmPACT DAB (Vector Laboratories) was used for the detection of horseradish peroxidase (HRP) activity. Slides were then dehydrated and mounted using SafeMount (Labonord).
Sánchez-Danés A., Larsimont J.C., Liagre M., Muñoz-Couselo E., Lapouge G., Brisebarre A., Dubois C., Suppa M., Sukumaran V., del Marmol V., Tabernero J, & Blanpain C. (2018). A slow-cycling Lgr5 tumour population mediates basal cell carcinoma relapse after therapy. Nature, 562(7727), 434-438.
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2

p53 Immunohistochemistry Protocol

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For p53 immunohistochemistry, 4-μm paraffin sections were deparaffinized, rehydrated, followed by antigen unmasking performed for 20 min at 98°C in citrate buffer (pH 6) using the PT module. Endogenous peroxydase was blocked using 3% H2O2 (Merck) in methanol for 10 min at room temperature. Endogenous avidin and biotin were blocked using the Endogenous Blocking kit (Invitrogen) for 20 min at room temperature. In p53 staining, nonspecific antigen blocking was performed using M.O.M. Basic kit reagent. Mouse anti-p53 antibody (clone 1C12; Cell Signaling) was incubated overnight at 4°C. Anti-mouse biotinylated in M.O.M. Blocking kit, Stan- dard ABC kit, and ImmPACT DAB (Vector Laboratories) was used for the detection of HRP activity. Slides were then dehydrated and mounted using SafeMount (Labonord).
Sánchez-Danés A., Hannezo E., Larsimont J.C., Liagre M., Youssef K.K., Simons B.D, & Blanpain C. (2016). Defining the clonal dynamics leading to mouse skin tumour initiation. Nature, 536(7616), 298-303.
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3

p53 Immunohistochemistry Protocol

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For p53 immunohistochemistry, 4-μm paraffin sections were deparaffinized, rehydrated, followed by antigen unmasking performed for 20 min at 98°C in citrate buffer (pH 6) using the PT module. Endogenous peroxydase was blocked using 3% H2O2 (Merck) in methanol for 10 min at room temperature. Endogenous avidin and biotin were blocked using the Endogenous Blocking kit (Invitrogen) for 20 min at room temperature. In p53 staining, nonspecific antigen blocking was performed using M.O.M. Basic kit reagent. Mouse anti-p53 antibody (clone 1C12; Cell Signaling) was incubated overnight at 4°C. Anti-mouse biotinylated in M.O.M. Blocking kit, Stan- dard ABC kit, and ImmPACT DAB (Vector Laboratories) was used for the detection of HRP activity. Slides were then dehydrated and mounted using SafeMount (Labonord).
Sánchez-Danés A., Hannezo E., Larsimont J.C., Liagre M., Youssef K.K., Simons B.D, & Blanpain C. (2016). Defining the clonal dynamics leading to mouse skin tumour initiation. Nature, 536(7616), 298-303.
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4

Immunohistochemical Analysis of Tissue Markers

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For staining on paraffin sections, 4μm paraffin sections were deparaffinized and rehydrated. Antigen unmasking was performed for 20 min at 98°C in citrate buffer (pH 6) using the PT module. Endogenous peroxidase was blocked using 3% H2O2 (Merck) in methanol for 10 min at RT. Endogenous avidin and biotin were blocked using the Endogenous Blocking kit (Invitrogen) for 20 min at RT. Nonspecific antigen blocking was performed using blocking buffer. Rabbit anti- YAP (1:200, Santa Cruz Biotechnology, sc-15407), rabbit anti- TAZ (1:100, Sigma- Aldrich, HPA007415), rabbit anti-cFOS (1:100, Proteintech, 26192-1-AP), rabbit anti-MYH9 (1:500, Sigma, HPA001644), rabbit anti-pERK (1:200, Cell signaling, 4370S) were incubated overnight at 4°C. For anti-FOSL1 (1:500, Santa Cruz Biotechnology, sc-28310) and anti-β-catenin (mouse, 1:1000 Abcam ab6301) tissue were also blocked with Mouse on Mouse (M.O.M.™) Blocking Reagent (Vector Laboratories). Anti- rabbit biotinylated with blocking buffer, anti-mouse biotinylated with blocking buffer, Standard ABC kit, and ImmPACT DAB (Vector Laboratories) were used for the detection of horseradish peroxidase (HRP) activity. Slides were then dehydrated and mounted using SafeMount (Labonord).
Aragona M., Sifrim A., Malfait M., Song Y., Van Herck J., Dekoninck S., Gargouri S., Lapouge G., Swedlund B., Dubois C., Baatsen P., Vints K., Han S., Tissir F., Voet T., Simons B.D, & Blanpain C. (2020). Mechanisms of stretch-mediated skin expansion at single-cell resolution. Nature, 584(7820), 268-273.
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5

Immunohistochemical Analysis of Key Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
For K14, Ki67, K10 and Lef1 immunohistochemistry in human samples, paraffin sections were deparaffinized, rehydrated, followed by antigen unmasking performed for 20 min at 98 °C in citrate buffer (pH 6) using the PT module. Endogenous peroxidase was blocked using 3% H2O2 (Merck) in methanol for 10 min at room temperature. Endogenous avidin and biotin were blocked using the Endogenous Blocking kit (Invitrogen) for 20 min at room temperature. Nonspecific antigen blocking was performed using blocking buffer. Mouse anti-K14 (rabbit, 1/2000, Thermofisher), anti-Ki67 (rabbit, 1/400, Abcam, ab15580), anti-Keratin10 (rabbit, 1/200, Biolegend, ref.90541) and anti-Lef1 (rabbit, 1/100, Cell Signaling, ref.2230) were incubated overnight at 4 °C. Anti-rabbit biotinylated with blocking buffer, Standard ABC kit, and ImmPACT DAB (Vector Laboratories) was used for the detection of horseradish peroxidase (HRP) activity. Slides were then dehydrated and mounted using SafeMount (Labonord).
Sánchez-Danés A., Larsimont J.C., Liagre M., Muñoz-Couselo E., Lapouge G., Brisebarre A., Dubois C., Suppa M., Sukumaran V., del Marmol V., Tabernero J, & Blanpain C. (2018). A slow-cycling Lgr5 tumour population mediates basal cell carcinoma relapse after therapy. Nature, 562(7727), 434-438.
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