Mice were sacrificed on indicated days. Tumor tissues were completely dissected, fixed in 4% buffered paraformaldehyde, embedded in paraffin, and were subsequently cut into 5-µm-thick sections. The sections were dewaxed and then placed in citric acid buffer (PH 6.0) for microwave antigen retrieval. Endogenous peroxidase of sections was then blocked with 3% hydrogen peroxide solution. Sections were further blocked with 3% bovine serum albumin blocking solution (Sigma) and immunostained with rabbit anti-mouse PD-L1 (ab233482), rabbit anti-mouse α-smooth muscle actin (SMA) (ab5694), rabbit anti-FN1 (ab2413), rabbit anti-CDH1 (ab76319) and rabbit anti-Ki-67 (ab15580) purchased from Abcam, UK at 4 ℃ overnight. The sections were then washed well with phosphate buffered saline with Tween-20 (PBST) and incubated with goat anti-rabbit IgG secondary antibody (ab150077, abcam) at room temperature for 1 hours. After being washed with PBST, staining was developed with DiAminoBenzidine. Slides were counterstained with hematoxylin, dehydrated, and finally mounted. These sections were observed under a microscope and were measured by using Image-pro Plus 6.0 software.
Bovine serum albumin blocking solution
Manufactured by Merck Group
Sourced in United Kingdom
Bovine serum albumin (BSA) blocking solution is a laboratory reagent used to prevent non-specific binding in various immunoassays and biochemical techniques. It contains a high concentration of purified BSA, which helps to block unoccupied binding sites on the test surface, reducing the likelihood of unwanted interactions and improving the specificity of the assay.
Automatically generated - may contain errors
Lab products found in correlation
Ab76319, by Abcam (1 mentions)
Ab233482, by Abcam (1 mentions)
Ab15580, by Abcam (1 mentions)
Ab2413, by Abcam (1 mentions)
Ab5694, by Abcam (1 mentions)
Ab150077, by Abcam (1 mentions)
Ab209538, by Abcam (1 mentions)
Ab150075, by Abcam (1 mentions)
Lsm 880 confocal microscope, by Zeiss (1 mentions)
2 protocols using bovine serum albumin blocking solution
1
Immunohistochemical Analysis of Tumor Microenvironment
2
Immunofluorescence Staining of Amyloid Aggregates
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The slides were fixed in 4% paraformaldehyde for 10 min and then washed with PBS for three times. After rinsing, slides were permeabilized in 1% Triton X-100 for 15 min. After three washings of 5 min each, slides were incubated in 5% Bovine serum albumin blocking solution (Sigma, Poole, United Kingdom) for 2 h. After blocking, the slides were incubated overnight with 0.05% Thioflavin T (ThT) and MJFR-14-6-4-2 (diluted 1:500, ab209538, Abcam, Cambridge, MA, United States). After overnight incubation, the slides were incubated with donkey anti-rabbit Alexa Fluor 647 (diluted 1:500, ab150075, Abcam, Cambridge, MA, United States) for 1 h at room temperature. Nuclei were stained with DAPI (0.2 μg/ml) for 5 min. Slides observation was performed with a Zeiss LSM 880 confocal microscope (Carl Zeiss Microscopy GmbH, Oberkochen, Germany) using 40 × objective.
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