Immunohistochemistry (IHC) staining and immunofluorescent (IF) staining were performed to examine and localize CD248 expression in RCC tissues and adjacent normal tissues. The primary antibodies used were as follows: CD248 (#ab204914, Abcam, Cambridge, UK), CD31 (#89C2, Cell Signaling Technology, USA), CD3 (#2100567, eBioscience, USA), CD206/MRC1 (#24595, Cell Signaling Technology, USA). The second antibodies were as follows: goat anti-rabbit immunoglobin [IgG; H&L; horse radish peroxidase (HRP); #ab6721, Abcam], donkey anti-rabbit IgG (#ab150076, Abcam), donkey anti-mouse IgG (#ab150105, Abcam). Nuclei were stained with DAPI (#C1002, Beyotime, Shanghai, China). Quantification was performed according to the percentage and intensity in IHC staining and the percentage of the positive area in the IF staining using Image J v1.52a (NIH, Bethesda, MD, USA).
Ab204914
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab204914 is an antibody-based product designed for laboratory use. It functions as a detection tool for the identification and characterization of target proteins or molecules in samples. The core purpose of this product is to provide researchers with a reliable and specific detection method for their experimental needs.
Automatically generated - may contain errors
Lab products found in correlation
Ab6721, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Ab150076, by Abcam (1 mentions)
Cd206 mrc1, by Cell Signaling Technology (1 mentions)
Ab150105, by Abcam (1 mentions)
Ab76104, by Abcam (1 mentions)
Fv 10 asw 1, by Olympus (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Fv1000, by Olympus (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Envision technique, by Agilent Technologies (1 mentions)
4 protocols using ab204914
1
Immunostaining of CD248 in Renal Cell Carcinoma
2
Triple Immunofluorescent Staining of PSMA, CD248, and CD31
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Triple immunofluorescent (IF) staining of PSMA, CD248, and CD31 was conducted for paraffin sections of UCB tissues. Briefly, the sections were incubated at room temperature with a mixture of anti-human CD248 (#ab204914, Abcam), anti-human PSMA (#ab76104, Abcam), and anti-human CD31 antibody (#3528, Cell Signaling Technology) for 16–18 h. The sections were then incubated with a mixture of Alexa488-conjugated donkey anti-goat IgG (#ab6721, Abcam), Cy3-conjugated donkey anti-rabbit IgG (#96907, Abcam), and Alexa647-conjugated donkey anti-mouse IgG (#ab150076, Abcam) for 4 h at room temperature. Nuclei were stained with 4'6-diamidino-2-phenylindole (DAPI) (#C1002, Beyotime, Jiangsu, China) or Hoechst 33342 (#C1022, Beyotime). The slides were covered, sealed with Vectashield (Vector, Burlingame, CA, USA), and observed under a confocal laser scanning microscope (FV1000; Olympus, Tokyo, Japan). The digital images were captured and processed using FV10-ASW 1.6 software (Olympus).
3
Immunohistochemical Analysis of EN and CD8
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4
Immunohistochemical Analysis of CD248
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Paraffin-embedded tissue microarrays (Outdo Biotech, Shanghai, China) were deparaffinized, rehydrated, and treated with 3% hydrogen peroxide for 10 min to inhibit endogenous peroxidase activity. Heat-mediated retrieval of antigens was performed in citrate buffer for 2 min. After being blocked with 5% bovine serum albumin (BSA) for 30 min, slides were incubated with rabbit anti-human CD248 primary antibody (1:2,000, ab204914, Abcam, MA, USA) overnight at 4°C. The immunodetection was performed using the standard rapid EnVision technique (Dako, Glostrup, Denmark). Subsequently, slides were washed in distilled water and counterstained with hematoxylin. Digital images for qualitative evaluation were obtained using an optical microscope (BX51, Olympus, Tokyo, Japan).
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