Anti rabbit igg horseradish peroxidase hrp

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-rabbit IgG-horseradish peroxidase (HRP) is a secondary antibody conjugate used in immunodetection applications. It binds to primary rabbit antibodies and includes the HRP enzyme, which can catalyze a colorimetric reaction for signal detection.

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Horseradish peroxidase (175 citations) Horseradish peroxidase (HRP) (40 citations) IgG rabbit (9 citations) Rabbit anti-IgG (8 citations) Anti-HRP rabbit (4 citations) Horseradish peroxidase anti-rabbit (3 citations) Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit and anti-mouse IgGs (2 citations)

6 protocols using anti rabbit igg horseradish peroxidase hrp

Immunohistochemical Analysis of BDNF and TrkB

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After the behavioral tests, mice were perfused with phosphate-buffered saline (PBS, pH 7.4) under inhaled CO2 anesthetization. The tumor tissues and brains were immediately removed from the mice and fixed in 4% paraformaldehyde for 48 h at 4 °C and transferred to 30% sucrose solutions for H&E and IHC staining. The mouse tissues frozen sections were blocked with 1% bovine serum albumin (BSA) diluted in PBS for 1 h at room temperature (RT); the sections were then blotted and incubated with rabbit recombinant antibody for BDNF (abcam, #ab108319) or rabbit polyclonal antibody for TrkB (abcam, #ab18987) at the appropriate dilution (1:40 dilution) in blocking serum for overnight at 4 °C. The slides were washed in PBS, followed by the anti-rabbit IgG-horseradish peroxidase (HRP) (1:500; Cell Signaling Technology, MA, USA #7074S) secondary antibody. The slides were washed, the peroxidase reaction was developed with diaminobenzidine and peroxide, and the slides were mounted in Aqua-Mount and evaluated on a light microscopy (Microscope Axio Imager.A2, Carl Zeiss, Oberkochen, Germany). A negative control was performed by omitting the primary antibody.

Yeo I.J., Yu J.E., Kim S.H., Kim D.H., Jo M., Son D.J., Yun J., Han S.B, & Hong J.T. (2024). TNF receptor 2 knockout mouse had reduced lung cancer growth and schizophrenia-like behavior through a decrease in TrkB-dependent BDNF level. Archives of Pharmacal Research, 47(4), 341-359.

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Immunoblotting of Raf-MEK-ERK Pathway

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Purified CD4⁺ T cells were suspended in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine and penicillin/streptomycin then left unstimulated or stimulated with 50 ng/ml PMA for 15 min at 37°C. Where indicated, cells were incubated in the presence or absence of Hyd (10 μM), H₂O₂ (50 μM), or ONOO⁻ (50 μM) for 60 min before PMA stimulation. Following stimulation the cells were lysed, then total and phosphorylated ERK, MEK, RAF and PKCδ measured by immunoblotting as described (16 (link)). The following primary Abs were used at a 1/1000 dilution: anti-phospho-raf (Ser³³⁸), anti-phospho-MEK1/2 (Ser^217/221), anti-MEK1/2 and anti-phospho-PKCδ (T⁵⁰⁵) (Cell Signaling Technology, Beverly MA). Polyclonal rabbit anti-active MAPK (1/5000) was purchased from Promega. Secondary Abs included anti-rabbit IgG horse radish peroxidase (HRP) (1/2000; Cell Signaling Technology) and anti-mouse IgG HRP (1/4000; Sigma-Aldrich).

Li Y., Gorelik G., Strickland F.M, & Richardson B.C. (2014). Oxidative Stress, T Cell DNA Methylation, and Lupus. Arthritis & Rheumatology (Hoboken, N.j.), 66(6), 1574-1582.

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Investigating MAPK Signaling Modulation

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Purified CD4+ T cells were suspended in RPMI 1640 supplemented with 10% fetal calf serum, 2 mM glutamine and penicillin/streptomycin and then left unstimulated or stimulated with 50 ng/ml phorbol myristate acetate (PMA) for 15 minutes at 37°C. Where indicated, cells were incubated in the presence or absence of hydralazine (10 μM), H₂O₂ (50 μM), or ONOO⁻ (50 μM) for 60 minutes before PMA stimulation. Following stimulation, the cells were lysed, then total and phosphorylated ERK, MEK, RAF, and PKCδ were measured by immunoblotting as described (16 (link)). The following primary antibodies were used at a 1:1,000 dilution: anti–phospho-Raf (Ser³³⁸), anti–phospho–MEK-1/2 (Ser^217/221), anti–MEK-1/2, and anti–phospho-PKCδ (T⁵⁰⁵) (Cell Signaling Technology). Polyclonal rabbit anti-active MAPK (1:5,000) was purchased from Promega. Secondary antibodies included anti-rabbit IgG horseradish peroxidase (HRP; 1:2,000) (Cell Signaling Technology) and anti-mouse IgG HRP (1:4,000; Sigma-Aldrich).

Li Y., Gorelik G., Strickland F.M, & Richardson B.C. (2014). Oxidative Stress, T Cell DNA Methylation, and Lupus. Arthritis & Rheumatology (Hoboken, N.j.), 66(6), 1574-1582.

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Western Blot Analysis of Osteoclastogenesis Regulators

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The reagents used were bought from the following specified manufacturers: rabbit monoclonal anti-c-fos, cleaved caspase 3, caspase 3, rabbit polyclonal anti-Pim-2, horseradish peroxidase (HRP)-anti-rabbit IgG, and anti-mouse IgG from Cell Signaling Technology (Beverly, MA, USA); mouse monoclonal anti-β-actin antibody from Sigma-Aldrich (St. Louis, MO, USA); mouse monoclonal antibodies against NFATc1 and CTSK from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA); human macrophage colony-stimulating factor (M-CSF) from Wako (Osaka, Japan); human soluble RANKL from Oriental Yeast Co. Ltd (Shiga, Japan); febuxostat from TEIJIN (Osaka, Japan); NAC from Nacalai tesqure (Kyoto, Japan); and Dox from Tokyo Chemical Industry (Tokyo, Japan).

Ashtar M., Tenshin H., Teramachi J., Bat-Erdene A., Hiasa M., Oda A., Tanimoto K., Shimizu S., Higa Y., Harada T., Oura M., Sogabe K., Nakamura S., Fujii S., Sumitani R., Miki H., Udaka K., Takahashi M., Kagawa K., Endo I., Tanaka E., Matsumoto T, & Abe M. (2020). The Roles of ROS Generation in RANKL-Induced Osteoclastogenesis: Suppressive Effects of Febuxostat. Cancers, 12(4), 929.

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Signaling Pathway Regulation in Cell Lines

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The following reagents were purchased from the indicated manufacturers: rabbit polyclonal anti-human DR4 antibody, anti-human TRPV1 antibody, and anti-human HDAC1 antibody from Abcam (Cambridge, UK); mouse polyclonal anti-β-actin antibody and valproate from Sigma (St. Louis, MO); rabbit polyclonal anti-Histone H3 antibody, rabbit polyclonal anti-acetyl-histone H3 antibody and anti-acetyl-histone H4 antibody from Merck Millipore (Billerica, MA); rabbit polyclonal anti-Sp1 antibody, anti-Akt antibody, anti-phosphorylated Akt antibody, anti-PI3K antibody, anti-phosphorylated PI3K antibody, anti-STAT3 antibody, anti-phosphorylated STAT3 antibody, anti-p38 antibody, anti-phosphorylated p38 antibody, anti-p65 antibody, anti-phosphorylated p65 antibody, anti-ERK antibody, anti-phosphorylated ERK antibody, anti-JNK antibody, anti-phosphorylated JNK antibody, horseradish peroxidase (HRP)-anti-rabbit IgG and anti mouse IgG from Cell Signaling Technology (Beverly, MA); rh IGF-1 from R&D Systems (Minneapolis, MN); panobinostat from Cayman Chemical Company (Ann Arbor, MI); and 5-azacytidine and Akt inhibitor VIII from Calbiochem (Darmstadt, Germany). The human monoclonal anti-DR4 agonistic IgG antibody R1-B12 was kindly provided by Kyowa Hakko Kirin Co. Ltd. (Tokyo, Japan).

Amachi R., Hiasa M., Teramachi J., Harada T., Oda A., Nakamura S., Hanson D., Watanabe K., Fujii S., Miki H., Kagawa K., Iwasa M., Endo I., Kondo T., Yoshida S., Aihara K.I., Kurahashi K., Kuroda Y., Horikawa H., Tanaka E., Matsumoto T, & Abe M. (2016). A vicious cycle between acid sensing and survival signaling in myeloma cells: acid-induced epigenetic alteration. Oncotarget, 7(43), 70447-70461.

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Inhibition of Apoptosis Pathways in Cancer Cells

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The following reagents purchased from the indicated manufacturers: rabbit polyclonal anti-human Sp1, IRF4, cleaved caspase-8 antibody, mouse polyclonal anti-human caspase-8 antibody, horseradish peroxidase (HRP)-anti-rabbit IgG, anti-mouse IgG and bortezomib from Cell Signaling Technology (Beverly, MA); rabbit polyclonal anti-human cMyc antibody and anti-HDAC1 antibody from Abcam (Cambridge, UK); anti-acetylated histone H3, acetylated histone H4 (Merk Millipore, Billerica, MA); mouse monoclonal anti-β-actin antibody and terameprocol from Sigma-Aldrich (St. Louis, MO); panobinostat from Cayman Chemical Company (Ann Arbor, MI); Z-lle-Glu(O-ME)-Thr-Asp(O-Me) fluoromethyl ketone (Z-IETD-FMK) from TONBO biosciences (San Diego, CA) and carfilzomib from Chemie Tek (Indianapolis, Indiana) respectively.

Bat-Erdene A., Miki H., Oda A., Nakamura S., Teramachi J., Amachi R., Tenshin H., Hiasa M., Iwasa M., Harada T., Fujii S., Sogabe K., Kagawa K., Yoshida S., Endo I., Aihara K, & Abe M. (2016). Synergistic targeting of Sp1, a critical transcription factor for myeloma cell growth and survival, by panobinostat and proteasome inhibitors. Oncotarget, 7(48), 79064-79075.

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