primary abs against gfp  (Cell Signaling Technology Inc)

Journal: Frontiers in Immunology

Article Title: A method for distinguishing and separating retinal microglia from infiltrating macrophages during an ocular inflammatory event

doi: 10.3389/fimmu.2026.1951506

Figure Lengend Snippet: Fluorescence microscopy of representative control (B6/J, Ccr2 RFP+/+ Cx3cr1 GFP+/+ , and R161H neg x Ccr2 RFP+/- Cx3cr1 GFP+/- F 1 ) and SAU (R161H +/- x Ccr2 RFP+/- Cx3cr1 GFP+/- F 1 ) mouse retinas for RFP and GFP at 50 days of age. 40x magnification.

Article Snippet: Primary antibodies against GFP (cat no. NB600-597, mouse anti-GFP, Novus Biologicals, Centennial, CO, USA) and RFP (cat no. NBP3-25634, rabbit anti-RFP, Novus Biologicals) were diluted 1:200 with DPBS and applied to the retinas and held at 4°C for at least 12 hours in dark conditions then washed 3 x 5 minutes with DPBS.

Techniques: Fluorescence, Microscopy, Control

Journal: Frontiers in Immunology

Article Title: A method for distinguishing and separating retinal microglia from infiltrating macrophages during an ocular inflammatory event

doi: 10.3389/fimmu.2026.1951506

Figure Lengend Snippet: Flow cytometry distinguishes retinal MG from infiltrating macrophages. (A, B) Analysis of peripheral blood mononuclear cells for RFP and GFP expression showing the background levels of RFP and GFP fluorescence in B6/J mice (A) and the presence of RFP + , GFP + and RFP + GFP + cells in the circulation of Ccr2 RFP+/+ Cx3cr1 GFP+/+ mice (B) . The gating strategy for selecting CD45 + cells from B6/J mice is the same as shown for Ccr2 RFP+/+ Cx3cr1 GFP+/+ mice. (C) Representative analysis of a B6/J mouse showing background levels of RFP and GFP fluorescence in retinal CD45 + cells. Background fluorescence for GFP indicated as GFP neg cells. (D) Representative analysis of R161H neg x Ccr2 RFP+/- Cx3cr1 GFP+/- F 1 retinas showing that a minimal number of all CD45 + cells (gate 1) express Ccr2 (RFP), and that a single population comprising most of the CD45 + cells and specifically the CD45 med cells (gate 2) express a high level of Cx3cr1 (GFP). Retinal CD45 + cells negative for GFP expression (GFP neg ) or with a low or intermediated level of GFP expression (GFP lo-med ) are indicated. (E) Representative analysis of R161H +/- x Ccr2 RFP+/- Cx3cr1 GFP+/- F 1 retinas with SAU showing a large increase in both CD45 med and CD45 hi cells with most of the RFP + cells found in the CD45 hi population as either RFP + GFP neg or RFP + GFP lo-med cells and that they are myeloid cells or lymphocytes. For all analyses; retinal MG indicated by red circles, flow cytometry done at 50 days of age, n = 4 for all groups of mice, and the mean percentage of cells in each quadrant for all mice analyzed is indicated.

Article Snippet: Primary antibodies against GFP (cat no. NB600-597, mouse anti-GFP, Novus Biologicals, Centennial, CO, USA) and RFP (cat no. NBP3-25634, rabbit anti-RFP, Novus Biologicals) were diluted 1:200 with DPBS and applied to the retinas and held at 4°C for at least 12 hours in dark conditions then washed 3 x 5 minutes with DPBS.

Techniques: Flow Cytometry, Analysis, Expressing, Fluorescence

Journal: Frontiers in Immunology

Article Title: A method for distinguishing and separating retinal microglia from infiltrating macrophages during an ocular inflammatory event

doi: 10.3389/fimmu.2026.1951506

Figure Lengend Snippet: Analysis of R161H neg x Ccr2 RFP+/- Cx3cr1 GFP+/- F 1 retinas for Ccr2 + (RFP + ) cells following ONC. (A) Representative fundoscopy of pre-ONC and day 10 post-ONC retinas. (B) Representative flow cytometry analysis of retinal CD45 med cells for RFP and GFP at day 10 post-ONC with mean percentage of CD45 med cells expressing GFP and RFP from all examined retinas indicated.

Article Snippet: Primary antibodies against GFP (cat no. NB600-597, mouse anti-GFP, Novus Biologicals, Centennial, CO, USA) and RFP (cat no. NBP3-25634, rabbit anti-RFP, Novus Biologicals) were diluted 1:200 with DPBS and applied to the retinas and held at 4°C for at least 12 hours in dark conditions then washed 3 x 5 minutes with DPBS.

Techniques: Analysis, Flow Cytometry, Expressing

Journal: bioRxiv

Article Title: Loss of Kmt2c / d promotes gastric cancer initiation and confers vulnerability to mTORC1 inhibition and anti-PD1 immunotherapy

doi: 10.1101/2025.03.27.645747

Figure Lengend Snippet: A, Schematic of mouse models: two doses of tamoxifen (3 mg × 2) were injected intraperitoneally with a 48-hour interval. B, Representative immunofluorescence (IF) staining of EYFP in stomach tissues from Tmprss2-CreER T2 ;Rosa26-LSL-EYFP mice . Nuclei were counterstained with DAPI. Scale bar, 200 µm. C, Kaplan-Meier plots showing the survival of mice after gene knockout. D, Stomach weight of the indicated genetically engineered mouse model (GEMM) six weeks after tamoxifen injection. Data are presented as mean ± SD and analyzed with two-tailed t-test. E, Thickness in corpus mucosa measured by microscopy of hematoxylin and eosin (H&E) staining section. Each dot represents the averaged thickness of 5 random fields from one mouse. Data are presented as mean ± SD and analyzed with two-tailed t-test. F, Representative H&E staining of stomach tissues after tamoxifen administration. Scale bar, 200 µm. G, Pathological staging of stomach cancer progression based on infiltration of tumor cells in TPCD mice. H, Representative H&E staining of small and large intestines in TPCD mice. Scale bar, 1 mm.

Article Snippet: Immunofluorescent staining was performed using primary antibodies against EYFP (2956, Cell Signaling Technology, 1:100), H3K4me1 (5326, Cell Signaling Technology, 1:100), ATP4A (D031-3, MBL Life Science, 1:500), and Puromycin (MABE343, Millipore Sigma, 1:100).

Techniques: Injection, Immunofluorescence, Staining, Gene Knockout, Two Tailed Test, Microscopy