Immunohistochemical Evaluation of Immune Markers

The paraffin sections were rehydrated with gradient ethanol solution. After antigen repair, the endogenous peroxidase activity was eliminated. The sections were blocked before incubation with the primary antibody (SLC1A5, abcom, ab237704; Anti -Iba1 Rabbit pAb, Servicebio, GB113502; Anti -CD86 Rabbit mAb, Servicebio, GB13585; Anti -CD163 Rabbit pAb, Servicebio, GB113152). SP Rabbit HRP Kit (DAB) (Cat.CW20355, CWBiotech) provides the protocol for the procedure. The results of IHC were scored according to the German Immunohistochemical Scores, and staining intensity at or above 4 points was defined as positive expression [47 (link)].

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Han L., Zhou J., Li L., Wu X., Shi Y., Cui W., Zhang S., Hu Q., Wang J., Bai H., Liu H., Guo C., Cao H., Chao M., Hu Y., Mou Y., Jiao Y., Feng D., Wang L, & Qu Y. (2022). SLC1A5 enhances malignant phenotypes through modulating ferroptosis status and immune microenvironment in glioma. Cell Death & Disease, 13(12), 1071.

Publication 2022

Anti -Iba1 Rabbit pAb

Antibody Antigen Cd163 Ethanol Paraffin Peroxidase Rabbit

Corresponding Organization :

Other organizations : Air Force Medical University

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Variable analysis

independent variables

Different primary antibodies used for immunohistochemistry (IHC): SLC1A5, Anti-Iba1, Anti-CD86, Anti-CD163

dependent variables

Expression levels of the target proteins (SLC1A5, Iba1, CD86, CD163) as determined by IHC staining

control variables

Gradient ethanol solution used for rehydration of paraffin sections
Antigen repair procedure
Elimination of endogenous peroxidase activity
Blocking of sections before incubation with primary antibodies
SP Rabbit HRP Kit (DAB) used for IHC staining protocol
German Immunohistochemical Scores used to evaluate and define positive expression (score ≥ 4)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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