Serial 5-µm paraffin sections were deparaffinized using xylene and serial ethanol dilutions and then stained with hematoxylin for 20 min and then eosin for 20 min at room temperature. Endogenous peroxidase was inactivated by treatment with 3% hydrogen peroxide solution for 10 min at room temperature. Sections were then washed with PBS, treated with 1% BSA for 30 min at room temperature and incubated with anti-CD44 (anti-homing receptor, cloneA020: MilliporeSigma) antibody at 1:200 dilution for 24 h at room temperature. Antigen retrieval for CMKLR1 antigen was performed using a microwave rapid sample processor MI-77 (Azumaya) with settings of 80˚C, 20 min and an output of 6. Sections were then incubated with anti-chemokine-like receptor 1 polyclonal antibody (anti-CMKLR1; Cayman Chemical Company) diluted at 1:100, followed by incubation with peroxidase-labelled goat anti-rat IgG (Bethyl Laboratories, Inc.) or Nichirei Histofine R (Nichirei Biosciences Inc.) diluted at 1:500 for 30 min. DAB staining was then performed and the nuclei were counterstained with hematoxylin for 1 min at room temperature.
Goat anti rat igg
Manufactured by Fortis Life Sciences
Sourced in United States
Goat anti-rat IgG is a laboratory reagent used for the detection and identification of rat immunoglobulin G (IgG) in biological samples. It is a polyclonal antibody produced by immunizing goats with rat IgG and purifying the specific antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Chemiluminescence detection reagent, by GE Healthcare (1 mentions)
Coomassie blue r 250, by Avantor (1 mentions)
Las 4000 image capture system, by Fujifilm (1 mentions)
Mouse anti his, by Qiagen (1 mentions)
Mouse anti gfp, by Takara Bio (1 mentions)
Mouse anti il 6, by Abcam (1 mentions)
Rat anti ha, by Roche (1 mentions)
2 protocols using goat anti rat igg
1
Immunohistochemical Staining of CD44 and CMKLR1
2
Protein Separation and Immunodetection
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TSP or purified proteins were separated using 10–12.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under the reducing condition, and the proteins on the gels were either stained with 0.25% Coomassie blue R-250 (AMRESCO, cat. no: 6104-59-2, Zottegem, Belgium) solution or analyzed by Western blotting using mouse anti-His (Qiagen, Valenica, CA, USA), mouse anti-LIF (abcam, cat. no., ab34427, Cambridge, UK), mouse anti-GFP (Clontech, cat. No., 632381, CA, USA), rat anti-HA (Roche, cat. No., 1583 816, Basel, Switzerland), mouse anti-IL6 (abcam, cat. no., ab9324, Cambridge, UK), and anti-CBD (Bioapp, Pohang, Korea) antibodies. The horseradish peroxidase (HRP)-conjugated sheep anti-mouse IgG (Bethyl, cat. no., A90-146P, TX, USA), and goat anti-rat IgG (Bethyl, cat. no., A110-105P, TX, USA) were used as secondary antibody. Bands on the immunoblots were detected using chemiluminescence detection reagents (GE healthcare, Illinois, USA), and images were captured with a LAS 4000 image capture system (Fujifilm, Tokyo, Japan).
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