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Biotinylated anti rabbit or polyvalent secondary antibody

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Biotinylated anti-rabbit or polyvalent secondary antibody is a laboratory reagent used for the detection and visualization of target proteins or antigens in various immunoassays. It binds to primary antibodies raised against rabbit or multiple species, and the biotin label enables signal amplification through the biotin-streptavidin interaction.

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Lab products found in correlation

Target retrieval buffer, by Vector Laboratories (5 mentions) Peroxidase blocking buffer, by Thermo Fisher Scientific (5 mentions) Protein block, by Thermo Fisher Scientific (5 mentions) S0809, by Agilent Technologies (4 mentions) Mouse on mouse m o m blocking reagent mkb 2213, by Vector Laboratories (2 mentions)

Spelling variants of this product

Anti-rabbit secondary antibody (58 citations) Biotinylated secondary antibody (32 citations) Biotinylated polyvalent secondary antibody (2 citations)

5 protocols using biotinylated anti rabbit or polyvalent secondary antibody

1

Immunohistochemistry Staining Protocol

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All antibodies are described in the Supplementary Information. Patient samples were collected under IRB exemption approval for protocol #EX21205258-1. Paraffin embedded sections were rehydrated through a xylene and alcohol series, rinsed in H2O and washed in PBS. Antigen retrieval was performed using target retrieval buffer (Vector Labs, Burlingame, CA) and steamed for 20 min. Samples were then blocked in a peroxidase blocking buffer (Thermo Scientific) for 15 min, followed by Protein block (Thermo Scientific) for 5 min, and incubated in appropriate primary antibody diluted in antibody diluent (S0809, Dako) at 4°C overnight in a humidified chamber. For mouse samples to be incubated with anti-mouse antibody, samples were blocked for 1 hour in Mouse on Mouse (M.O.M.™) Blocking Reagent (MKB-2213, Vector Labs). Following washing in PBS, samples were incubated in biotinylated anti-rabbit or polyvalent secondary antibody (Thermo Scientific) followed by streptavidin-HRP solution at room temperature for 20 min. Samples were then washed in PBS and incubated in 3-Amino-9-Ethyl-l-Carboazole (AEC) chromogen and counter stained with Mayers hematoxylin for 1 min, rinsed in cold H2O, and mounted in Aquamount.
Kaur A., Webster M.R., Marchbank K., Behera R., Ndoye A., Kugel CH I.I.I., Dang V.M., Appleton J., O’Connell M.P., Cheng P., Valiga A.A., Morissette R., McDonnell N.B., Ferrucci L., Kossenkov A.V., Meeth K., Tang H.Y., Yin X., Wood WH I.I.I., Lehrmann E., Becker K.G., Flaherty K.T., Frederick D.T., Wargo J.A., Cooper Z.A., Tetzlaff M.T., Hudgens C., Aird K.M., Zhang R., Xu X., Liu Q., Bartlett E., Karakousis G., Eroglu Z., Lo R.S., Chan M., Menzies A.M., Long G.V., Johnson D.B., Sosman J., Schilling B., Schadendorf D., Speicher D.W., Bosenberg M., Ribas A, & Weeraratna A.T. (2016). sFRP2 in the aged microenvironment drives melanoma metastasis and therapy resistance. Nature, 532(7598), 250-254.
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2

Paraffin-Embedded Immunohistochemistry Protocol

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Immunohistochemistry was performed on paraffin embedded sections as
described previously (18 ). Briefly,
Paraffin embedded sections were rehydrated through a xylene and alcohol series
rinsed in H2O and washed in PBS. Antigen retrieval was performed by steaming for
20 min in target retrieval buffer (Vector Labs, Burlingame, CA) and steamed for
20 min. Samples were blocked in a peroxidase blocking buffer (Thermo Scientific)
followed by Protein block (Thermo Scientific) and incubated in appropriate
primary antibody diluted in antibody diluent (S0809, Dako) at 4°C
overnight in a humidified chamber. Samples were then incubated in biotinylated
anti-rabbit or polyvalent secondary antibody (Thermo Scientific) followed by
streptavidin-HRP solution. Samples were then washed in PBS and incubated in
3-Amino-9-Ethyl-l-Carboazole (AEC) chromogen. Finally, samples were washed in
H2O, incubated in Meyer’s hematoxylin for 1 min, rinsed in cold H2O, and
mounted in Aquamount. Patient samples were collected under IRB exemption
approval for protocol #EX21205258-1.
Behera R., Kaur A., Webster M.R., Kim S., Ndoye A., Kugel CH I.I.I., Alicea G.M., Wang J., Ghosh K., Cheng P., Lisanti S., Marchbank K., Dang V., Levesque M., Dummer R., Xu X., Herlyn M., Aplin A.E., Roesch A., Caino C., Altieri D.C, & Weeraratna A.T. (2017). Inhibition of age-related therapy resistance in melanoma by rosiglitazone-mediated induction of Klotho. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(12), 3181-3190.
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3

Immunohistochemistry Staining Protocol

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All antibodies are described in the Supplementary Information. Patient samples were collected under IRB exemption approval for protocol #EX21205258-1. Paraffin embedded sections were rehydrated through a xylene and alcohol series, rinsed in H2O and washed in PBS. Antigen retrieval was performed using target retrieval buffer (Vector Labs, Burlingame, CA) and steamed for 20 min. Samples were then blocked in a peroxidase blocking buffer (Thermo Scientific) for 15 min, followed by Protein block (Thermo Scientific) for 5 min, and incubated in appropriate primary antibody diluted in antibody diluent (S0809, Dako) at 4°C overnight in a humidified chamber. For mouse samples to be incubated with anti-mouse antibody, samples were blocked for 1 hour in Mouse on Mouse (M.O.M.™) Blocking Reagent (MKB-2213, Vector Labs). Following washing in PBS, samples were incubated in biotinylated anti-rabbit or polyvalent secondary antibody (Thermo Scientific) followed by streptavidin-HRP solution at room temperature for 20 min. Samples were then washed in PBS and incubated in 3-Amino-9-Ethyl-l-Carboazole (AEC) chromogen and counter stained with Mayers hematoxylin for 1 min, rinsed in cold H2O, and mounted in Aquamount.
Kaur A., Webster M.R., Marchbank K., Behera R., Ndoye A., Kugel CH I.I.I., Dang V.M., Appleton J., O’Connell M.P., Cheng P., Valiga A.A., Morissette R., McDonnell N.B., Ferrucci L., Kossenkov A.V., Meeth K., Tang H.Y., Yin X., Wood WH I.I.I., Lehrmann E., Becker K.G., Flaherty K.T., Frederick D.T., Wargo J.A., Cooper Z.A., Tetzlaff M.T., Hudgens C., Aird K.M., Zhang R., Xu X., Liu Q., Bartlett E., Karakousis G., Eroglu Z., Lo R.S., Chan M., Menzies A.M., Long G.V., Johnson D.B., Sosman J., Schilling B., Schadendorf D., Speicher D.W., Bosenberg M., Ribas A, & Weeraratna A.T. (2016). sFRP2 in the aged microenvironment drives melanoma metastasis and therapy resistance. Nature, 532(7598), 250-254.
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4

Immunohistochemical Staining of Mouse Tumors

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Mouse tumors were paraffin embedded and sectioned. Paraffin embedded sections were rehydrated through a xylene and alcohol series, rinsed in H2O and washed in PBS. The slides were then fixed with 10% formalin for 10 min and washed in PBS. Antigen retrieval was performed using target retrieval buffer (Vector Labs, Burlingame, CA) and steamed for 20 min. Samples were then blocked in a peroxidase blocking buffer (Thermo Scientific) for 15 min, followed by Protein block (Thermo Scientific) for 5 min, and incubated in appropriate primary antibody diluted in antibody diluent (S0809, Dako) at 4°C overnight in a humidified chamber. Following washing in PBS, samples were incubated in biotinylated anti-rabbit or polyvalent secondary antibody (Thermo Scientific) followed by streptavidin-HRP solution at room temperature for 20 min. Samples were then washed in PBS and incubated in 3-Amino-9-Ethyl-l-Carboazole (AEC) chromogen for up to 10 min. Finally, samples were washed in H2O, incubated in Mayers hematoxylin for 1 min, rinsed in cold H2O, and mounted in Aquamount.
Webster M.R., Fane M.E., Alicea G.M., Basu S., Kossenkov A.V., Marino G.E., Douglass S.M., Kaur A., Ecker B.L., Gnanapradeepan K., Ndoye A., Kugel C., Valiga A., Palmer J., Liu Q., Xu X., Morris J., Yin X., Wu H., Xu W., Zheng C., Karakousis G.C., Amaravadi R.K., Mitchell T.C., Almeida F.V., Xiao M., Rebecca V.W., Wang Y.J., Schuchter L.M., Herlyn M., Murphy M.E, & Weeraratna A.T. (2019). Paradoxical role for wild type p53 in driving therapy resistance in melanoma. Molecular cell, 77(3), 633-644.e5.
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5

Immunohistochemical Analysis of Mouse Tumors

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Mouse tumors were paraffin embedded and sectioned. Paraffin embedded sections were rehydrated through a xylene and alcohol series, rinsed in H 2 O and washed in PBS. The slides were then fixed with 10% formalin for 10 min and washed in PBS. Antigen retrieval was performed using target retrieval buffer (Vector Labs, Burlingame, CA) and steamed for 20 min. Samples were then blocked in a peroxidase blocking buffer (Thermo Scientific) for 15 min, followed by Protein block (Thermo Scientific) for 5 min, and incubated in appropriate primary antibody diluted in antibody diluent (S0809, Dako) at 4°C overnight in a humidified chamber. Following washing in PBS, samples were incubated in biotinylated anti-rabbit or polyvalent secondary antibody (Thermo Scientific) followed by streptavidin-HRP solution at room temperature for 20 min. Samples were then washed in PBS and incubated in 3-Amino-9-Ethyl-l-Carboazole (AEC) chromogen for up to 10 min. Finally, samples were washed in H 2 O, incubated in Mayers hematoxylin for 1 min, rinsed in cold H 2 O, and mounted in Aquamount.
Webster M.R., Fane M.E., Alicea G.M., Basu S., Kossenkov A.V., Marino G.E., Douglass S.M., Kaur A., Ecker B.L., Gnanapradeepan K., Ndoye A., Kugel C.H., Valiga A.A., Palmer J., Liu Q., Xu X., Morris J., Yin X., Wu H., Xu W., Zheng C., Karakousis G.C., Amaravadi R.K., Mitchell T.C., Almeida F.V., Xiao M., Rebecca V.W., Wang Y.J., Schuchter L.M., Herlyn M., Murphy M.E., & Weeraratna A.T. (2020). Paradoxical Role for Wild-Type p53 in Driving Therapy Resistance in Melanoma. Molecular Cell, 77(3), 681-681.
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