Paraffin-embedded lung sections (4-µm thickness) were used for this study. Images were taken under polarized light using an upright dry 40× objective. Tissues were deparaffinized and after antigen retrieval, permeabilized with 0.4% triton X-100. Tissues were blocked with 10% BSA and maintained in PBS + 5% BSA + 0.4% triton X-100 throughout antibody treatments. Primary antibodies were incubated at 4°C overnight and secondary antibodies were incubated for 1 hr at room temperature. Primary antibodies used (1:200 dilution) include rabbit anti-NFκB p65 (#8242, Cells Signaling Technology); mouse anti-cRel (#MA5-15859, Thermo Fisher Scientific); rabbit anti-NFκB1 (#13586, Cells Signaling Technology); mouse anti-NFκB1 (#NBP2-66976, Novus); rabbit anti-RUNX1(#ab229482, Abcam); goat anti-IL33 (#AF3626 R&D); mouse anti-ICAM1 (#sc-1511, Santa Cruz Biotechnology, Inc); and mouse anti-CD3 ((# sc-7296, Santa Cruz Biotechnology, Inc). Goat anti-rabbit and anti-mouse IgG-A594 or IgG-A488 were used as secondary antibodies (1:200 dilution at RT). For anti-goat, rabbit anti-goat IgGA-647 was used as secondary antibody (1:200 dilution at RT). DAPI was used for nuclear counterstaining. The ProLong Gold antifade reagent was used as mounting media. Mount slides were examined with a Leica DM6000 B. Slide Book 6 (3i) was used for analysis and capturing images.
Mouse anti cd3
Manufactured by Santa Cruz Biotechnology
Sourced in China
Mouse anti-CD3 is an antibody that recognizes the CD3 antigen, which is a protein complex found on the surface of T cells. This antibody can be used for the identification and isolation of T cells in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Dm6000b, by Leica (1 mentions)
Rabbit anti runx1, by Abcam (1 mentions)
Goat anti il33, by R&D Systems (1 mentions)
Mouse anti icam 1, by Santa Cruz Biotechnology (1 mentions)
Mouse anti crel, by Thermo Fisher Scientific (1 mentions)
Rabbit anti cd11b, by Novus Biologicals (1 mentions)
Rabbit anti nf κb p65, by Cell Signaling Technology (1 mentions)
Rabbit anti fhl2, by Abcam (1 mentions)
Rabbit anti phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Dab kit, by Vector Laboratories (1 mentions)
Dab kit, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Rabbit anti zo 1, by Abcam (1 mentions)
4 protocols using mouse anti cd3
1
Immunohistochemical Analysis of Lung Tissue
2
Tumor Microvasculature and Immune Cells
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See Table S3 . Tumor microvascular density was assessed using the rat anti-mouse CD31 (PECAM-1, clone SZ31) antibody (Dianova). The following antibodies were used to characterize immune cell subpopulations in the LL/2 tumors: rat anti-CD45 (Novus Biologicals), rabbit anti-CD11 b, rat anti-Ly6G/Ly6c (Novusbio), mouse anti-CD3, rat anti-CD4, goat anti-CD8 (Santa Cruz).
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3
Immunohistochemical Analysis of Lung Tissue
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Paraffin-embedded lung sections (4-µm thickness) were used for this study. Images were taken under polarized light using an upright dry under 40× objective. Tissues were deparaffinized and, after antigen retrieval, permeabilized with 0.4% Triton X-100. Tissues were blocked with 10% BSA and maintained in PBS + 5% BSA + 0.4% Triton X-100 throughout antibody treatments. Primary antibodies were incubated at 4°C overnight, and secondary antibodies were incubated for 1 h at room temperature. Primary antibodies used (1:200 dilution) included rabbit anti-phospho-p44/42 MAPK (Erk1/2; Thr202/Tyr204; #9101S; Cell Signaling Technology); rabbit anti-NF-κB p65 (#8242; Cell Signaling Technology); mouse anti-NFATC2 (#sc-7296, Santa Cruz Biotechnology) mouse anti-CD3 (#sc-7296; Santa Cruz Biotechnology); rabbit anti-Fhl2 (#ab12328; Abcam); and mouse anti-PAR2 (SAM11, #817201; BioLegend). Goat anti-rabbit IgG-A594 or IgG-A488 was used as secondary antibody (1:200 dilution at room temperature). DAPI was used for nuclear counterstaining. ProLong Gold antifade reagent was used as mounting media. Mounted slides were examined with a Leica DM6000 B. Slide Book 6 (3i) was used for analysis and capturing images.
4
Immunohistochemical Analysis of Intestinal Tight Junctions and T-Cell Markers
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Paraffin-embedded intestinal tissue sections were used for immunohistochemistry. Tight junction proteins ZO-1 and Occludin expression was detected using rabbit anti-ZO-1 (1:1000, Abcam, Cambridge, MA, USA) and rabbit anti-Occludin antibodies (1:50, Abcam, Cambridge, MA, USA), respectively. Thereafter, the sections were immunostained with goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Zhongshan Goldenbridge, Beijing, China).
T-cell markers CD3, CD4, CD8a, and CD25 were detected using mouse anti-CD3, rat anti-CD4, mouse anti-CD8a and mouse anti-CD25 (1:250, Santa Cruz Biotechnologies, Dallas, TX, USA), respectively. For the secondary antibody, we utilized goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Vector Laboratories, Inc., Burlingame, CA, USA).
T-cell markers CD3, CD4, CD8a, and CD25 were detected using mouse anti-CD3, rat anti-CD4, mouse anti-CD8a and mouse anti-CD25 (1:250, Santa Cruz Biotechnologies, Dallas, TX, USA), respectively. For the secondary antibody, we utilized goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP) using a DAB kit (Vector Laboratories, Inc., Burlingame, CA, USA).
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