RBL-2H3 cells (1 x 105) were cultured on glass coverslips, sensitized with anti-DNP IgE (1 μg/ml, 16 h) and stimulated with antigen (DNP-BSA, 30 ng/ml). The cells were fixed with 4% paraformaldehyde, and permeabilized with 0.5% saponin for 10 min. The slides were incubated with PBS containing 1% BSA at room temperature for 1 h, washed with PBS with 0.1% Tween (PBST) and incubated with anti-NHERF1 antibody (Abcam, 1:500 dilution) at room temperature for 1 h. Cells were washed with PBST and incubated with Alexa Fluor 555-conjugated anti-rabbit IgG secondary antibody (Invitrogen, 1:300 dilution) for 1 h at room temperature in the dark followed by incubation in 300 μM DAPI (1:1000 dilution). Slides were washed and mounted using the ProLong™ Diamond antifade mounting media (Invitrogen). Confocal images were obtained with the Olympus FV 1000 confocal laser scanning microscope (Olympus America, Center Valley, PA) with a 40X objective and images were analyzed using the ImageJ software.
Anti nherf1 antibody
Manufactured by Abcam
Sourced in United States
Anti-NHERF1 antibody is a laboratory reagent used for the detection and analysis of NHERF1 (Na+/H+ exchanger regulatory factor 1) protein in biological samples. NHERF1 is a scaffolding protein that plays a role in the regulation of various cellular processes. This antibody can be used in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to identify and study the NHERF1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Prolong diamond antifade mounting media, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555 conjugated anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal laser scanning microscope, by Olympus (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Ventana benchmark gx, by Roche (1 mentions)
2 protocols using anti nherf1 antibody
1
Visualizing NHERF1 Translocation in Stimulated RBL-2H3 Cells
2
Quantitative Analysis of NHERF1 in Lung Tissue
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The 5 μm sections of collected lung tissues were deparaffinized, rehydrated, rinsed with distilled water, and washed with Tris-buffered saline (TBS). Automated IHC staining was carried out using a Ventana BenchMark GX instrument (Ventana Medical Systems, Inc., Tucson, AZ, USA). NHERF1 was probed with an anti-NHERF1 antibody (1:100, Abcam, Cambridge, MA, USA). Image Pro Plus 6.0 (Media Cybernetics, Inc., Rockville, MD, USA) was used to calculate the intensity of NHERF1 staining. Randomly selected 10 microscopic fields were imaged for each sample subjected to computer-assisted imaging analyses. The results were presented as the mean ± standard deviation (SD). The determination of immunohistochemical staining intensity and positive areas for NHERF1 in IOD (integral optical density) measures were performed by two independent staffs who were mutually blinded to the sample grouping information.
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