To confirm foamy macrophages by staining (Figure S3K ), 4 μm tissue sections of CRC patients 380 and 393 were deparaffinized, rehydrated, stained with H&E, and imaged at 20× magnification. Subsequently, slide coverslips were removed, and antigen retrieval was performed in EDTA pH 9 buffer for 5 min in 95 °C in a pressure cooker. Slides were next stained with CD68 XP monoclonal antibody (1/200, rabbit, D4B9C, Cell Signaling Technology Cat# 76437, RRID:AB_2799882) and imaged at 20× magnification.
D4b9c
Manufactured by Cell Signaling Technology
Sourced in United States
D4B9C is a high-quality primary antibody for use in various immunoassays. It specifically recognizes and binds to its target antigen, enabling detection and analysis of the corresponding protein.
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Lab products found in correlation
3 protocols using d4b9c
1
Confirming Foamy Macrophages in CRC Patients
2
Multiplex Immunohistochemistry Antibodies
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Monoclonal rabbit anti-human PD-L1 antibody (E1L3N, #13684) and monoclonal rabbit anti-human CD68 antibody (D4B9C, #76437) were obtained from Cell Signaling Technology and the polyclonal rabbit anti-human/mouse CD47 antibody (ab175388), monoclonal rabbit anti-human/mouse CD163 antibody (clone EPR19518), monoclonal rabbit anti-mouse PD-L1 antibody (clone EPR20529), rabbit anti-CD4 antibody (EPR19514), anti-CD8 antibody (YTS169.4), rabbit anti-iNOS antibody (ab15323), and rabbit Anti-CD206 antibody (ab64693) were from Abcam. Anti-mouse CD8a monoclonal antibody, PE (Clone: 53-6.7) were purchased from eBioscience. Anti-mouse CD279 (PD-1), FITC (Clone: 29F.1A12) were purchased from Biolegend.
3
Quantifying Tumor-Associated Macrophages in Tissues
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IHC staining was used to evaluate the intratumoral and peritumoral presence of TAMs (CD68-positive) and M2-like macrophages (CD163-positive), a subtype of TAM, in primary and metastasis site tissues. Five micrometer-thick sequential tissue sections were obtained from paraffin-embedded tissue samples and used for the IHC analysis as described in previous studies[15 (link),16 (link)]. The primary antibodies used in IHC staining were antibodies against CD68 (rabbit anti-human, D4B9C, dilution 1:400; Cell Signaling Technology, Danvers, MA, United States) and CD163 (rabbit anti-human, D6U1J, dilution 1:500; Cell Signaling Technology). Anti-rabbit IgG, horseradish peroxidase-linked antibody was used as the secondary antibody. For the assessment of expression of CD68- and CD163-positive cells, the tissue sections were screened using each immunohistochemistry slide at the low power fields (× 4), and the hot spots were selected. Immune cell staining was scored by counting the number of stained immune cells in three high power fields (× 400) in the hot spots. IHC sections were independently assessed by three authors (Yang J, Chen YH, and Zhou L).
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