Mice were anesthetized by overdose with pentobarbital and perfused transcardially with PBS followed by 4% paraformaldehyde in 1X PBS. DRGs were dissected and fixed in 4% PFA for 2 h, followed by 30% sucrose with 0.02% azide in 1X PBS. DRGs were placed in OCT and cut on a cryostat at 10 μm, followed by mounting on SuperFrost Plus glass slides for staining. Infiltration of neutrophils into the DRG neurons in naïve, control, and primed mice was evaluated using immunohistochemical staining of S100A9 costained with NeuN, a marker for neurons. The tissue sections were blocked with 10% normal goat serum (Vector Labs S-1000) in PBSTx (0.3% Triton-X in 1×PBS). Sections were costained with a monoclonal rat anti-S100A9 (Abcam; ab105472) and recombinant rabbit anti-NeuN antibody (Abcam; ab177487) and their respective secondary antibodies (Alexa Fluor 594 anti-rabbit IgG, ThermoFisher; A11037 and Alexa Fluor 488 goat anti-rat, Abcam; ab150157). Sections were mounted and coverslipped with Vectashield Hardset Antifade Mounting Medium with DAPI (Vectorlabs; H-1500) and imaged using an Olympus fluorescent microscope. The total number of S100A9+ cell neurons was quantified in three DRG sections per slide and pooled per mouse using ImageJ. Neutrophils within 250 μm of the tissue periphery were counted by a blinded experimenter using ImageJ.
A11037
Manufactured by Abcam
Sourced in United Kingdom
A11037 is a laboratory equipment product. It serves as a core function without further interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Ab199428, by Abcam (2 mentions)
Formalin, by Thermo Fisher Scientific (2 mentions)
Ab90996, by Abcam (2 mentions)
Ab95997, by Abcam (2 mentions)
Ab17721, by Abcam (2 mentions)
Ab150157, by Abcam (1 mentions)
Ab105472, by Abcam (1 mentions)
Ab177487, by Abcam (1 mentions)
S 1000, by Vector Laboratories (1 mentions)
Alexa fluor 594 anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
H1500, by Olympus (1 mentions)
3 protocols using a11037
1
Quantification of Neutrophil Infiltration in Dorsal Root Ganglia
2
Immunofluorescence Staining of Cancer Cells
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MCF7 and T47D cells were seeded on slides at an appropriate density for 24h. After paraformaldehyde fixation, cells on slides were treated with 0.3% Triton X-100 for permeabilization of the plasma membrane and then immunostained with specific antibodies including anti-LSD1 (Abcam, ab90996 1:200 or ab17721; 1:200), anti-GATA3 (Abcam, ab199428; 1:200), anti-TRIM37 (Abcam, ab95997; 1:200), and DAPI (nuclear marker). The expression of target proteins (red or green) and DAPI (blue) were examined by fluorescence microscopy; areas of co-localization are shown in the merged images.
For the mouse model, IF staining was performed on lung tissue sections derived from the indicated mouse models injected with Ad-K8-Cre that were fixed in 10% formalin (Fisher Scientific, Hampton, NH) and embedded in paraffin. Antigen retrieval (Citrate buffer pH 6.0, 20 min boil in microwave oven, level 1) was performed prior to blocking. Primary antibody [anti-KDM1/LSD1 (ab17721, 1:500, Abcam, Cambridge, UK)] and the secondary antibody [goat anti-rabbit IgG conjugated with AF594 (A11037, 1:250)] were used. Slides were counterstained with DAPI (1 μg/ml).
For the mouse model, IF staining was performed on lung tissue sections derived from the indicated mouse models injected with Ad-K8-Cre that were fixed in 10% formalin (Fisher Scientific, Hampton, NH) and embedded in paraffin. Antigen retrieval (Citrate buffer pH 6.0, 20 min boil in microwave oven, level 1) was performed prior to blocking. Primary antibody [anti-KDM1/LSD1 (ab17721, 1:500, Abcam, Cambridge, UK)] and the secondary antibody [goat anti-rabbit IgG conjugated with AF594 (A11037, 1:250)] were used. Slides were counterstained with DAPI (1 μg/ml).
3
Immunofluorescence Staining of Cancer Cells
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