Five-μm-thick ice sections were stained with hematoxylin and eosin staining or subjected to immunohistology with an anti-α smooth muscle actin (α-SMA) antibody (ab5694, Abcam Japan, Tokyo, Japan), an anti-neutrophil antibody (ab2557, Abcam Japan, Tokyo, Japan), and an anti-Mac-3 antibody (550292, BD Pharmingen, Tokyo, Japan). To detect primary antibodies, sections for the anti-α-SMA antibody were incubated with the Dako Envision+ system HRP-labeled polymer anti-rabbit (ready to use) (Dako North America, California, USA) and sections for the anti-neutrophil and anti-Mac-3 antibodies were incubated with polyclonal rabbit anti-rat immunoglobulins/HRP (Dako North America, California, USA). These immunohistological staining was performed in accordance with our previous studies [21 (link),25 (link),30 (link),31 (link)]. Negative control slides were obtained by omitting each primary antibody.
Polyclonal rabbit anti rat immunoglobulins hrp
Manufactured by Agilent Technologies
Sourced in United States, Japan
Polyclonal rabbit anti-rat immunoglobulins/HRP is a laboratory reagent used for the detection and quantification of rat immunoglobulins in various immunoassay techniques. It consists of polyclonal antibodies raised in rabbits against rat immunoglobulins, which are conjugated with horseradish peroxidase (HRP) enzyme for signal amplification.
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Lab products found in correlation
Ab2557, by Abcam (1 mentions)
Dako envision system hrp labeled polymer anti rabbit, by Agilent Technologies (1 mentions)
Ab5694, by Abcam (1 mentions)
Polyclonal goat anti rabbit immunoglobulins hrp, by Agilent Technologies (1 mentions)
Liquid dab substrate chromogen system, by Agilent Technologies (1 mentions)
2 protocols using polyclonal rabbit anti rat immunoglobulins hrp
1
Immunohistological Analysis of Tissue Sections
2
Immunohistochemical Analysis of Extracellular Matrix
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Immunohistochemical staining was performed on paraffin sections. Endogenous peroxidase was blocked by immersing the sections in 0.3% H2O2 in methanol for 30 min at room temperature (RT). The specimens were blocked with 3% skim milk in 0.01% Tween-20-PBS for 60 min at RT and were then incubated overnight with a rat monoclonal primary antibody against TNC (1:100, Santa Cruz Biotechnology, Dallas, TX, USA), a rabbit polyclonal primary antibody against FN (1:200, Thermo Scientific, West Palm Beach, FL, USA) and a rabbit polyclonal primary antibody against α-SMA (1:200, GeneTex, Inc., Irvine, CA, USA). After washing with Tween-20-PBS, the specimens were treated with the appropriate secondary antibodies (polyclonal rabbit anti-rat immunoglobulins/HRP or polyclonal goat anti-rabbit immunoglobulins/HRP; Dako, Tokyo, Japan) for 60 min at RT. Color was developed using the Liquid DAB+Substrate Chromogen System (Dako) followed by counterstaining with Mayer’s haematoxylin (Muto Pure Chemicals, Tokyo, Japan).
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