Excised tumors from mice were fixed with 10% neutral buffered formalin for 24 hours and changed to 70% ethanol. Tumors were paraffin-embedded and sectioned by the Brain Tumor Research Center at UCSF. For IHC, slides were deparaffinized and antigen retrieval was performed using a pressure cooker. The Mouse on Mouse (M.O.M.™) basic kit (Vector laboratories) was used for masking endogenous mouse antigen. The VECTASTAIN® ABC reagent (Vector laboratories) was used for signal detection. The CD45 antibody (05–1410) was purchased from EMD Millipore and was used at a concentration of 1:50. The CD38 (ab216343), CD68 (ab955) and TMEM119 (ab209064) antibodies were purchased from Abcam and were used at a concentration of 1:100. The CD206 antibody (AF2535SP) was purchased from R&D Systems and was used at a concentration of 1:50. Images were taken using a Zeiss AxioImager M1 microscope. Cells were quantified using Image J.
Tmem119 ab209064
Manufactured by Abcam
TMEM119 (ab209064) is a recombinant protein that corresponds to the extracellular domain of the transmembrane protein TMEM119. TMEM119 is a microglial-specific marker that is expressed on the cell surface of microglia, the resident macrophages of the central nervous system. This protein can be used for the detection and identification of microglia in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Vectastain abc reagent, by Vector Laboratories (1 mentions)
Mouse on mouse m o m basic kit, by Vector Laboratories (1 mentions)
Axio imager m1 microscope, by Zeiss (1 mentions)
Mab382, by Merck Group (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Dig rna labeling kit, by Roche (1 mentions)
Image pro plus, by Media Cybernetics (1 mentions)
2 protocols using tmem119 ab209064
1
Immunohistochemical Analysis of Tumor Infiltrates
2
GAS5 RNA In Situ Hybridization and Immunocytochemistry
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An approximately 500‐bp mouse or human GAS5 sequence (Appendix Table S1 ) was cloned and ligated into the pEASY‐T5 plasmid (Transgen, Beijing, China) containing the T3 and T7 promoters, respectively. Antisense and sense digoxigenin‐labeled cRNA probes were synthesized with a DIG‐RNA Labeling Kit (12430721, Roche) with linearized templates. In situ hybridization was performed according to the previous report 43 .
For ICF, cells or tissue sections were fixed, permeabilized, and incubated with primary antibodies (MBP, MAB382, Chemicon; MAP2, M9942, Sigma‐Aldrich; TNF‐α, AF‐510‐NA, R&D; Arg‐1, 610708, BD; Olig2, AB9610, Millipore; CC1, ab16794, Abcam; TMEM119, ab209064, Abcam; and Iba1, 019‐19471, Wako) overnight at 4°C, followed by 2 h of incubation with TRITC‐ or FITC‐conjugated secondary antibodies (Jackson Immuno Research). Then, the samples were counterstained with Hoechst 33342 (Sigma‐Aldrich). Fluorescence images were captured using fluorescence microscopy (DXM, Nikon) or confocal microscopy (Leica) and analyzed using Image‐Pro Plus (Media Cybernetics).
For ICF, cells or tissue sections were fixed, permeabilized, and incubated with primary antibodies (MBP, MAB382, Chemicon; MAP2, M9942, Sigma‐Aldrich; TNF‐α, AF‐510‐NA, R&D; Arg‐1, 610708, BD; Olig2, AB9610, Millipore; CC1, ab16794, Abcam; TMEM119, ab209064, Abcam; and Iba1, 019‐19471, Wako) overnight at 4°C, followed by 2 h of incubation with TRITC‐ or FITC‐conjugated secondary antibodies (Jackson Immuno Research). Then, the samples were counterstained with Hoechst 33342 (Sigma‐Aldrich). Fluorescence images were captured using fluorescence microscopy (DXM, Nikon) or confocal microscopy (Leica) and analyzed using Image‐Pro Plus (Media Cybernetics).
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