Formalin-fixed paraffin-embedded (FFPE) blocks of GD thyroid tissues (n=4) were cut into 4-μm serial sections for immunohistochemical staining. Immunostaining for human CD11c (at a 1:300 dilution, Abcam, Cambridge, UK), human CD19 (at a 1:400 dilution, Abcam, Cambridge, UK), or CXCL10 (at a 1:200 dilution, Proteintech, China) was performed by 3,3’-diaminobenzidine (DAB) staining (Zhongshan Jinqiao Biotechnology, Beijing, China) according to the manufacturer’s instructions. CD19 antigen repair was conducted using citric acid solution (pH=6.0) for 15 min by microwave heating. CD11c and CXCL10 antigen repair was performed with Tris–EDTA buffer and heated for 15 min in a microwave. Two sections were prepared as negative controls for each immunostaining batch.
Cxcl10
Manufactured by Proteintech
Sourced in United Kingdom
CXCL10 is a chemokine protein that is involved in the recruitment and activation of immune cells. It plays a role in inflammatory and immune responses. This product is a high-quality recombinant protein for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Human cd11c, by Abcam (1 mentions)
Occludin, by Proteintech (1 mentions)
Cd206 mrc1, by Cell Signaling Technology (1 mentions)
Claudin 1, by Proteintech (1 mentions)
Tnf α, by BioLegend (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by BioLegend (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Tsg101, by GeneTex (1 mentions)
Sting, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Pano multiplex ihc kit, by Panovue (1 mentions)
Hif 1α, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
α sma, by Proteintech (1 mentions)
4 protocols using cxcl10
1
Immunohistochemical Analysis of Thyroid Tissues
2
Comprehensive Immunological Assay Protocol
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ELISA, immunoblot, and mRNA analysis were conducted as described [62 (link), 63 (link)]. ELISA kits included IFN-γ (Invitrogen, 88–8314-22), IL-1β (eBioscience, 88,701,388), IL-18 (MBL, D042-3), IL-6 (BioLegend, 431,301), and TNF-α (BioLegend, 430,901). Primary antibodies used in this study include antibodies against ALIX (Cell Signaling, 2171, 1:1000), TSG101 (Genetex, GTX70255; 1:500), GAPDH (Cell Signaling, 2118; 1:1000), CD206/MRC1 (Cell Signaling, 24,595; 1:1000), occludin (ProteinTech, 27,260-1-AP; 1:1000), claudin1 (ProteinTech, 13,050-1-AP; 1:1000), CD81 (ProteinTech, 27,855-1-AP; 1:1000), Cxcl10 (ProteinTech, 10,937-1-AP; 1:1000), Arg1 (ProteinTech, 66,129-1; 1:1000), FMOD (ProteinTech, 60,108-1; 1:500), and MFGE8 (ThermoFisher, PA5-109,955; 1:1000).
3
Immunohistochemical Analysis of Tumor Markers
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Immunohistochemistry was performed as previously described29 ,31 , on formalin-fixed paraffin-embedded tumor sections (5 μm) using primary antibodies at a 1:50–500 dilution following the manufacturers’ instructions directed against STING (#13647; Cell signaling), Ki-67 (#RB-9043-P0; Thermo Fisher Scientific), cleaved caspase-3 (#9664; Cell Signaling), IFNA1 (#18013-1-AP, Proteintech), IFNB (#27506-1-AP, Proteintech), CXCL10 (#10937-1-AP, Proteintech), and MX1 (#13750-1-AP, Proteintech). The staining was achieved by incubating with diaminobenzidine (DAB) for 5 min using a DAB peroxidase substrate kit (Vector Laboratories, Burlingame, CA, USA). IHC analysis was performed in triplicates for this study. On each slide, cell counting was conducted at 40X magnification, focusing on five randomly selected fields with a minimum count of 500 cells. Cells showing negative or positive staining by antibodies were counted separately. The percentage of cells exhibiting positive staining was calculated using the following formula: [(number of positively stained cells divided by the sum of negatively and positively stained cells) × 100%].
4
Multiplex Immunohistochemistry for Tumor Microenvironment
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Multiplex immunohistochemistry (mIHC) was performed using the PANO Multiplex IHC kit (Panovue, 10144100100). Formalin-fixed paraffin-embedded sections (3.5 µm) were deparaffinized and rehydrated. Each slide underwent several cycles of staining, including heat-induced epitope retrieval with citrate buffer (pH=6.0) or Tris/EDTA (pH=9.0), endogenous peroxidase blocking with 3% H2O2, and non-specific protein blocking with 10% goat serum, followed by incubation of primary antibodies and corresponding horseradish peroxidase-conjugated secondary antibody (Panovue, 10013001040). Finally, tyramide signal amplification dyes were applied to amplify fluorescence signals. The following primary antibodies were used in sequential rounds of staining: HIF-1α (SANTA CRUZ, Cat#sc-13515, 1:200), CD8α (CST, Cat#98941, 1:500), PD-1 (CST, Cat#84651, 1:400), CD31 (CST, Cat#77699, 1:500), α-SMA (ProteinTech, Cat#14 395–1-AP, 1:12000), Ki67 (Abcam, Cat#ab16667,1:200), GZMB (CST, Cat#44153, 1:200), CXCR3 (Abcam, Cat#ab288437, 1:500), CXCL10 (ProteinTech, Cat#10 937–1-AP, 1:500). Slides were then counterstained with DAPI (Beyotime, C1005) and mounted with antifade mountant (Panovue, 10022001010).
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