Hrp conjugated donkey anti rabbit secondary antibodies

Manufactured by Abcam

HRP-conjugated donkey anti-rabbit secondary antibodies are laboratory reagents used to detect the presence of rabbit primary antibodies in various immunoassays. These antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that catalyzes a colorimetric reaction, enabling the visualization and quantification of target proteins or antigens.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (2 mentions) Ab216347, by Abcam (1 mentions) Ab243136, by Abcam (1 mentions) Ab179467, by Abcam (1 mentions) Ecl system, by Thermo Fisher Scientific (1 mentions) Bca kit, by Beyotime (1 mentions) Pvdf membrane, by Merck Group (1 mentions)

Close spelling variants of this product

Rabbit antibodies (7 citations) Rabbit anti- (5 citations) Donkey anti-rabbit-HRP (4 citations) HRP-conjugated antibodies (4 citations) Anti-rabbit antibodies (4 citations) Anti-rabbit HRP secondary antibodies (2 citations)

3 protocols using hrp conjugated donkey anti rabbit secondary antibodies

Immunohistochemical Quantification of CCL13 and KLRC1

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The protein expression levels of CCL13 and KLRC1 were estimated via immunohistochemical (IHC) staining. Briefly, the tissues slides were deparaffinized, rehydrated and treated with 3% H₂O₂ for 15 min to eliminate endogenous peroxidase. Then, antigen retrieval was performed by heating the slides in sodium citrate buffer for 3 min. Next, the slides were incubated with rabbit anti-CCL13 or anti-KLRC1 primary antibodies (Affinity, Biosciences, 1:200) at 4°C overnight. The slides were washed and incubated with HRP-conjugated donkey anti-rabbit secondary antibodies (Abcam) for 15 min. The staining was visualized using DAB solution and samples were counterstained with hematoxylin.
Immunostaining of CCL13 and KLRC1 were analyzed by two pathologists who were blinded to the same information. The staining intensity score was defined on a scale of 0 to 3 in which 0 means no staining, 1 means mild staining, 2 means medium staining and 3 means intense staining. The percentage score of stained cells were also calculated on a scale of 1 to 4 in which 1 represents (0–25%), 2 = (26–50%), 3 = (51–75%) and 4 = (76–100%). In order to obtain the final score, the intensity score and percentage score were multiplied to reach the final score ranging from 0 to 12.

Liu J., Wang Y., Mei J., Nie S, & Zhang Y. (2021). Identification of a Novel Immune Landscape Signature for Predicting Prognosis and Response of Endometrial Carcinoma to Immunotherapy and Chemotherapy. Frontiers in Cell and Developmental Biology, 9, 671736.

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Western Blot Analysis of Liver Cancer Cell Proteins

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Treated liver cancer cells were lysed in RIPA buffer (cat no. 89900; Thermo Fisher Scientific, Inc.) for total protein extraction. The protein concentration was determined using a BCA kit (Beyotime Institute of Biotechnology) and the protein samples (40 µg) in each lane were separated with 10% SDS-PAGE. The target proteins were transferred to a PVDF membrane (EMD Millipore), which was incubated in 5% non-fat milk at room temperature for 90 min to block non-specific binding sites. Subsequently, the membranes were incubated at 4°C overnight with primary antibodies against pro-survival protein myeloid cell leukemia 1 (MCL1; rat product code ab243136; 1:1,000), Snail1 (rabbit; 1:1,000; product code ab216347) and actin (rabbit; product code ab179467; 1:5,000; all from Abcam). The membranes were probed with HRP-conjugated donkey anti-rabbit secondary antibodies (1:2,000; product code ab6802; Abcam) at room temperature for 1 h. The signals were visualized using an ECL system (Thermo Fisher Scientific, Inc.).

Yang G., Li X., Liu J., Huang S., Weng Y., Zhu J., Lin D, & Jiang O. (2021). Hsa_circ_0008537 facilitates liver carcinogenesis by upregulating MCL1 and Snail1 expression via miR-153-3p. Oncology Reports, 45(3), 1072-1082.

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Immunohistochemical Analysis of CCL13 and KLRC1

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The protein expression levels of CCL13 and KLRC1 were tested via immunohistochemical (IHC) staining. Briefly, the tissues slides were deparaffinized, rehydrated and treated with 3% H2O2 for 15 minutes to eliminate endogenous peroxidase. Then, antigen retrieval was performed by heating the slides in sodium citrate buffer for 3 minutes. Next, the slides were incubated with rabbit anti-CCL13 or anti-KLRC1 primary antibodies (Affinity, Biosciences, 1:200) at 4℃ overnight. The slides were washed and incubated with HRP-conjugated donkey anti-rabbit secondary antibodies (Abcam) for 15 minutes. The staining was visualized using DAB solution and samples were counterstained with hematoxylin.
Immunostaining of CCL13 and KLRC1 were analyzed by two pathologists who were blinded to the same information. The staining intensity score was defined on a scale of 0 to 3 in which 0 means no staining, 1 means mild staining, 2 means medium staining and 3 means intense staining. The percentage score of stained cells were also calculated on a scale of 1 to 4 in which 1 represents (0-25%), 2 = (26%-50%), 3 = (51%-75%) and 4 = (76%-100%). In order to obtain the final score, the intensity score and percentage score were multiplied to reach the final score ranging from 0 to 12.

Liu J., Wang Y., Mei J., Nie S., & Zhang Y. (2020). Identification of a Novel Immune Landscape Signature for Predicting Prognosis and Response of Endometrial Carcinoma to Immunotherapy and Chemotherapy.

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