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Hrp labeled goat anti rat igg h l

Manufactured by Beyotime
Sourced in China

HRP-labeled goat anti-rat IgG (H + L) is a laboratory reagent used for immunodetection and quantification applications. It is composed of polyclonal goat antibodies that bind to rat immunoglobulin G (IgG) molecules, both the heavy and light chains. These antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that can be used for colorimetric or chemiluminescent detection.

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3 protocols using hrp labeled goat anti rat igg h l

1

Protein-Protein Interaction Study via Co-Immunoprecipitation

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For transient transfection, 293T cells were co-transfected with HA-FOXR2/SRF and Myc-EPC1/EPC2 expression vectors. At 36 to 48 h post transfection, cells were collected for immunoprecipitation assays. The primary antibodies anti-Myc-Tag, anti-HA-Tag and anti-β-Actin, and secondary antibodies HRP-labeled goat anti-rabbit IgG (H + L) (Cat# A0208, Beyotime) and HRP-labeled goat anti-rat IgG (H + L) (Cat# A0192, Beyotime) were used. Anti-HA-Tag antibody-conjugated agarose beads were purchased from Sigma (Cat# A2095). After addition of cell extracts (transfected with Myc tagged EPC1/EPC2 and HA tagged FOXR2/SRF) and incubation for 4 h at 4°C, the bound beads were washed with PBS and analyzed by WB analysis. The Amersham Imager 600 analyzer was used for photographing the blots. ImageJ software was used for quantifying the protein levels based on the band density obtained in the WB analysis.
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2

Polyclonal Antibody Generation against rCgDM9CP-4

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The purified rCgDM9CP-4 was immuned to 6 weeks old rats with a weight of approximately 120 g to acquire polyclonal antibody. Briefly, rCgDM9CP-4 of 1mg mL-1 was mixed with freund’s adjuvant to immunize the female rats for four times.
After SDS-PAGE, the samples of rCgDM9CP-4 were electrophoretically transferred onto a nitrocellulose membrane. The membrane was blocked with PBST containing 5% milk at room temperature for 1 h, and incubated with antibodies (diluted at 1:4000) for 1 h at room temperature with rocking, followed by three times of washes with PBST. Membranes were incubated with HRP-labeled Goat Anti-Rat IgG(H+L) (Beyotime; 1:10000) for 1 h at room temperature with rocking, followed by three times of washes with PBST. Membranes were incubated briefly in Western lighting ECL substrate system (Thermo Scientific, USA) before exposure to KODAK X-OMAT AR X-ray film (Eastman Kodak, USA).
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3

Tissue Histology Analysis by H&E, Iron, and IHC

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For Hematoxylin and Eosin (H&E) staining, tissues were fixed for 24 h in 10% neutral buffered formalin, dehydrated and embedded in paraffin. Tissue sections (3–5 µm) were stained with H&E. Images were acquired with KFBIO KF-PRO-120 digital pathology slide scanner.
The frozen tissues were sliced into sections of 10 μm each at −20 °C and fixed for 10 min in 4% paraformaldehyde. Tissue sections were staining with Perls’ Iron Stain Kit (Solarbio, Beijing, China) following manufacturer’s instructions. When indicated, Perls’ staining was further enhanced using the DAB-kit (Beyotime, Shanghai, China). For immunohistochemistry, tissue sections were fixed, washed in PBS and treated with H2O2 to block endogenous peroxidase. Sections were incubated with anti-F4/80 (clone BM8, Biolegend, San Diego, CA, USA, 1:500) at 4 ℃ overnight and were incubated with HRP-labeled Goat Anti-Rat IgG (H + L) (Beyotime, Shanghai, China, 1:100) at 37 °C for 1 h and then stained with diaminobenzidine (DAB) at room temperature for 10 min in the dark, followed by counterstaining with hematoxylin for cell nuclei. Images were acquired with a Leica microscope.
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