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Beyotime peroxidase conjugated secondary antibody
Peroxidase Conjugated Secondary Antibody, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/peroxidase conjugated secondary antibody/product/Beyotime
Average 92 stars, based on 88 article reviews
Price from $9.99 to $1999.99
peroxidase conjugated secondary antibody - by Bioz Stars, 2020-07
92/100 stars

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Incubation:

Article Title: Astragalus membranaceus and Panax notoginseng, the Novel Renoprotective Compound, Synergistically Protect against Podocyte Injury in Streptozotocin-Induced Diabetic Rats
Article Snippet: .. The membrane was incubated overnight at 4°C with a primary antibody, then washed with TBST for five times and incubated with horseradish peroxidase-conjugated secondary antibodies (Beyotime, China) at room temperature for 1 hour and developed with an enhanced chemiluminescence agent. .. The membranes were placed on the ChemiDoc Touch Imaging System (Bio-Rad Laboratories, USA) to image a protein band, and ImageJ (Adobe Corp., USA) was used to determine band intensity.

Article Title: AhR-E2F1-KGFR signaling is involved in KGF-induced intestinal epithelial cell proliferation
Article Snippet: .. Membranes were blocked in 5% skim milk at 4°C for 1 h and incubated with anti-AhR (1:500 dilution), anti-KGFR (1:200 dilution), anti-E2F1 (1:1,000 dilution) or anti-GAPDH (1:1,000 dilution) antibodies for 24 h at 4°C, followed by incubation for 1 h with the peroxidase-conjugated secondary antibody (1:5,000 dilution; cat no. BA1055/BA1051; Beyotime Institute of Biotechnology). .. Protein bands were detected using a Kodak Gel Logic 4000R Imaging System (Kodak, Rochester, NY, USA) with an enhanced chemiluminescence reagent (Boster Systems Inc., Pleasanton, CA, USA).

Article Title: Arsenic trioxide induces apoptosis and the formation of reactive oxygen species in rat glioma cells
Article Snippet: .. Then, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies, and detected using an enhanced chemiluminescence (ECL) kit (Beyotime). ..

Article Title: Long Non-coding RNA LINC00339 Stimulates Glioma Vasculogenic Mimicry Formation by Regulating the miR-539-5p/TWIST1/MMPs Axis
Article Snippet: .. The membranes were blocked with blocking buffer (5% non-fat milk in 1% Tween Tris-buffered saline [TBS]) for 2 hr at room temperature, followed by incubation with primary antibodies overnight at 4°C, incubated with horseradish peroxidase-conjugated secondary antibodies for 2 hr at room temperature and then developed with the enhanced chemiluminescence reagents (Beyotime Institute of Biotechnology, China) according to the manufacturer’s protocol. .. For protein band analyses, an internal control GAPDH (Proteintech, USA) was used.

Article Title: IL13Rα2 siRNA inhibited cell proliferation, induced cell apoptosis, and suppressed cell invasion in papillary thyroid carcinoma cells
Article Snippet: .. Then the membrane was washed three times with Tris-buffered saline and incubated with horseradish peroxidase-conjugated secondary antibodies (Beyotime) for 1 hour. .. Immunoreactive bands were detected using an ECL detection kit (Millipore).

Article Title: miRNA-133b targets FGFR1 and presents multiple tumor suppressor activities in osteosarcoma
Article Snippet: .. After three washes with TBST, the membrane was incubated with horseradish peroxidase-conjugated secondary antibodies at room temperature for 2 h. The signal was developed using the ECL system (Beyotime, Jiangsu, China) according to the manufacturer’s instructions. ..

Article Title: Knockdown of SIRT7 enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells partly via activation of the Wnt/β-catenin signaling pathway
Article Snippet: .. After washing in TBST four times (5 min each), the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (anti-mouse or anti-rabbit; Beyotime) for 1 h at room temperature. ..

Article Title: Oxidative stress, autophagy and pyroptosis in the neovascularization of oxygen-induced retinopathy in mice
Article Snippet: .. The blots were subsequently incubated with horseradish peroxidase-conjugated secondary antibody (cat. no. A0208; 1:1,000; Beyotime Institute of Biotechnology) at room temperature for 2 h. All blots were developed using a chemiluminescence system (Fluor Chem M System; ProteinSimple, San Jose, CA, USA) and signal intensities were analyzed with a Gel-pro 4.5 Analyzer (Media Cybernetics, Inc., Rockville, MD, USA). ..

Blocking Assay:

Article Title: Long Non-coding RNA LINC00339 Stimulates Glioma Vasculogenic Mimicry Formation by Regulating the miR-539-5p/TWIST1/MMPs Axis
Article Snippet: .. The membranes were blocked with blocking buffer (5% non-fat milk in 1% Tween Tris-buffered saline [TBS]) for 2 hr at room temperature, followed by incubation with primary antibodies overnight at 4°C, incubated with horseradish peroxidase-conjugated secondary antibodies for 2 hr at room temperature and then developed with the enhanced chemiluminescence reagents (Beyotime Institute of Biotechnology, China) according to the manufacturer’s protocol. .. For protein band analyses, an internal control GAPDH (Proteintech, USA) was used.

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  • 92
    Beyotime hrp conjugated goat anti rabbit
    Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit or anti-mouse <t>IgG</t> (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.
    Hrp Conjugated Goat Anti Rabbit, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit/product/Beyotime
    Average 92 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti rabbit - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    92
    Beyotime peroxidase conjugated secondary antibody
    Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit or anti-mouse <t>IgG</t> (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.
    Peroxidase Conjugated Secondary Antibody, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase conjugated secondary antibody/product/Beyotime
    Average 92 stars, based on 88 article reviews
    Price from $9.99 to $1999.99
    peroxidase conjugated secondary antibody - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    89
    Beyotime anti rabbit mouse horseradish peroxidase conjugated secondary antibody
    Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit or anti-mouse <t>IgG</t> (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.
    Anti Rabbit Mouse Horseradish Peroxidase Conjugated Secondary Antibody, supplied by Beyotime, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit mouse horseradish peroxidase conjugated secondary antibody/product/Beyotime
    Average 89 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti rabbit mouse horseradish peroxidase conjugated secondary antibody - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    91
    Beyotime hrp conjugated goat anti rabbit igg secondary antibody
    Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit or anti-mouse <t>IgG</t> (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.
    Hrp Conjugated Goat Anti Rabbit Igg Secondary Antibody, supplied by Beyotime, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit igg secondary antibody/product/Beyotime
    Average 91 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti rabbit igg secondary antibody - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

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    Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase (HRP)-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.

    Journal: Frontiers in Endocrinology

    Article Title: A Type IIb, but Not Type IIa, GnRH Receptor Mediates GnRH-Induced Release of Growth Hormone in the Ricefield Eel

    doi: 10.3389/fendo.2018.00721

    Figure Lengend Snippet: Specificities of anti-GnRHR1 and anti-GnRHR2 antisera against recombinant proteins as determined by Western blot analysis. The recombinant proteins (50 ng) or COS-7 cell extracts (100 μg) were separated on 12% SDS-PAGE gels, processed routinely, and immnoreacted with the rabbit anti-GnRHR1 antiserum [1:2,000 dilution; (A) ], the mouse anti-GnRHR2 antiserum [1:1,000 dilution; (B) ], the anti-GnRHR1 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (C) , or the anti-GnRHR2 antiserum pre-adsorbed with 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) . The secondary antibody was 1:1,000 diluted horseradish peroxidase (HRP)-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (Beyotime, Shanghai, China), and the blots were visualized using BeyoECL Plus kit (Beyotime). GnRHR1-antigen, the recombinant GnRHR1 polypeptide used to immunize rabbit; GnRHR1-TRX, the region corresponding to GnRHR1 antigen expressed as thioredoxin (TRX) fusion protein; GnRHR2-TRX, GnRHR2 polypeptide region encompassing the synthetic antigen peptide expressed as TRX fusion protein. GnRHR1 and GnRHR2, the recombinant full-length ricefield eel GnRHR1 and GnRHR2 proteins expressed in transiently transfected COS-7 cells. pcDNA, extracts of COS-7 cells transfected with the empty vector pcDNA3.0. COS-7: extracts of COS-7 cells.

    Article Snippet: Then slides were applied to the secondary antibody, HRP-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (1:1,000 dilution; Beyotime), and finally visualized with 3,3'-diaminobenzidine (DAB) solution, mounted, and digitally photographed with a microscope (Eclipse Ni-U, Nikon, Japan).

    Techniques: Recombinant, Western Blot, SDS Page, Transfection, Plasmid Preparation

    Specificities of GnRHR1 and GnRHR2 immunoreactivities in the pituitary of male ricefield eels as determined by immunohistochemical analysis. Sagittal sections of the pituitary gland were immunoreacted with the rabbit anti-GnRHR1 antiserum [1:500 dilution; (A) ], the pre-adsorbed anti-GnRHR1 antiserum by 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (B) , the mouse anti-GnRHR2 antiserum [1:500 dilution; (C) ], the pre-adsorbed anti-GnRHR2 antiserum by 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) , or the mixture of the rabbit anti-GnRHR1 (1:500 dilution) with the mouse anti-GnRHR2 (1:500 dilution) antisera (E,F) . The secondary antibody was 1:1,000 diluted horseradish peroxidase (HRP)-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (Beyotime, Shanghai, China) for (A–D) , and the mixture of Alexa Flour 488-labeled goat anti-rabbit IgG (H+L) (1:500 dilution) and Cy3-labeled goat anti-mouse IgG (H+L) (1:500 dilution) for (E) . DAPI was used to stain the nuclei blue. Immunostaining (brown) in (A–D) was visualized by the DAB chromogen and counterstained with hematoxylin. The black triangles and stars indicate the adenohypophysis and neurohypophysis tissues, respectively. The image of (E) was captured and analyzed with a Nikon i-E confocal microscope equipped with a CSU-W1 spinning-disk head (Yokogawa, Tokyo, Japan) for immunofluorescent staining of GnRHR1 (green) and GnRHR2 (red). (F) is the higher magnification of the boxed area in (E) . Sagittal sections of ricefield eel pituitary glands were shown here with the rostral (anterior) to the left. Scale bar = 50 μm except the image of (F) was 10 μm.

    Journal: Frontiers in Endocrinology

    Article Title: A Type IIb, but Not Type IIa, GnRH Receptor Mediates GnRH-Induced Release of Growth Hormone in the Ricefield Eel

    doi: 10.3389/fendo.2018.00721

    Figure Lengend Snippet: Specificities of GnRHR1 and GnRHR2 immunoreactivities in the pituitary of male ricefield eels as determined by immunohistochemical analysis. Sagittal sections of the pituitary gland were immunoreacted with the rabbit anti-GnRHR1 antiserum [1:500 dilution; (A) ], the pre-adsorbed anti-GnRHR1 antiserum by 10 ug/mL of recombinant GnRHR1 expressed in transfected COS-7 cells (B) , the mouse anti-GnRHR2 antiserum [1:500 dilution; (C) ], the pre-adsorbed anti-GnRHR2 antiserum by 10 ug/mL of recombinant GnRHR2 expressed in transfected COS-7 cells (D) , or the mixture of the rabbit anti-GnRHR1 (1:500 dilution) with the mouse anti-GnRHR2 (1:500 dilution) antisera (E,F) . The secondary antibody was 1:1,000 diluted horseradish peroxidase (HRP)-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (Beyotime, Shanghai, China) for (A–D) , and the mixture of Alexa Flour 488-labeled goat anti-rabbit IgG (H+L) (1:500 dilution) and Cy3-labeled goat anti-mouse IgG (H+L) (1:500 dilution) for (E) . DAPI was used to stain the nuclei blue. Immunostaining (brown) in (A–D) was visualized by the DAB chromogen and counterstained with hematoxylin. The black triangles and stars indicate the adenohypophysis and neurohypophysis tissues, respectively. The image of (E) was captured and analyzed with a Nikon i-E confocal microscope equipped with a CSU-W1 spinning-disk head (Yokogawa, Tokyo, Japan) for immunofluorescent staining of GnRHR1 (green) and GnRHR2 (red). (F) is the higher magnification of the boxed area in (E) . Sagittal sections of ricefield eel pituitary glands were shown here with the rostral (anterior) to the left. Scale bar = 50 μm except the image of (F) was 10 μm.

    Article Snippet: Then slides were applied to the secondary antibody, HRP-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (1:1,000 dilution; Beyotime), and finally visualized with 3,3'-diaminobenzidine (DAB) solution, mounted, and digitally photographed with a microscope (Eclipse Ni-U, Nikon, Japan).

    Techniques: Immunohistochemistry, Recombinant, Transfection, Labeling, Staining, Immunostaining, Microscopy

    GnRH1, GnRH2, and GnRH3 immunostaining in the brain and pituitary of ricefield eels. Sagittal sections of the brains together with the pituitary gland of ricefield eels were immunoreacted with primary antibodies, the rabbit polyclonal antibody AS-691 for GnRH1 [1:7,000 dilution; (B,C,H) ], the rabbit polyclonal antibody 675 for GnRH2 [1:2,000 dilution; (D,I) ], or the mouse monoclonal antibody LRH13 for GnRH3 [1:2,000 dilution; (E–G,J) ]. After incubation with primary antibodies for 40 h at 4°C, sections were then exposed to the secondary antibody, HRP-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (1:1,000 dilution; Beyotime, Shanghai, China), and finally visualized with 3,3'-diaminobenzidine (DAB) solution, mounted, and digitally photographed with a Nikon Eclipse Ni-U microscope (Japan). (A) , the schematic diagram of the ricefield eel brain and pituitary. Red, green, and blue dots in (A) represent GnRH1, GnRH2, and GnRH3 immunoreactive neurons, respectively. OB, olfactory bulbs; TEL, telencephalon; VT, ventral telencephalon; POA, preoptic area; OT, optic tectum-thalamus; MT, midbrain tegmentum; Hyp, hypothalamus; Pit, pituitary; Ce, cerebellum; MO, medulla oblongata. Scale bar = 50 μm except where it is specifically designated otherwise.

    Journal: Frontiers in Endocrinology

    Article Title: A Type IIb, but Not Type IIa, GnRH Receptor Mediates GnRH-Induced Release of Growth Hormone in the Ricefield Eel

    doi: 10.3389/fendo.2018.00721

    Figure Lengend Snippet: GnRH1, GnRH2, and GnRH3 immunostaining in the brain and pituitary of ricefield eels. Sagittal sections of the brains together with the pituitary gland of ricefield eels were immunoreacted with primary antibodies, the rabbit polyclonal antibody AS-691 for GnRH1 [1:7,000 dilution; (B,C,H) ], the rabbit polyclonal antibody 675 for GnRH2 [1:2,000 dilution; (D,I) ], or the mouse monoclonal antibody LRH13 for GnRH3 [1:2,000 dilution; (E–G,J) ]. After incubation with primary antibodies for 40 h at 4°C, sections were then exposed to the secondary antibody, HRP-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (1:1,000 dilution; Beyotime, Shanghai, China), and finally visualized with 3,3'-diaminobenzidine (DAB) solution, mounted, and digitally photographed with a Nikon Eclipse Ni-U microscope (Japan). (A) , the schematic diagram of the ricefield eel brain and pituitary. Red, green, and blue dots in (A) represent GnRH1, GnRH2, and GnRH3 immunoreactive neurons, respectively. OB, olfactory bulbs; TEL, telencephalon; VT, ventral telencephalon; POA, preoptic area; OT, optic tectum-thalamus; MT, midbrain tegmentum; Hyp, hypothalamus; Pit, pituitary; Ce, cerebellum; MO, medulla oblongata. Scale bar = 50 μm except where it is specifically designated otherwise.

    Article Snippet: Then slides were applied to the secondary antibody, HRP-conjugated goat anti-rabbit or anti-mouse IgG (H+L) (1:1,000 dilution; Beyotime), and finally visualized with 3,3'-diaminobenzidine (DAB) solution, mounted, and digitally photographed with a microscope (Eclipse Ni-U, Nikon, Japan).

    Techniques: Immunostaining, Incubation, Microscopy