Formalin fixed Paraffin embedded H&E stained sections of cystectomy were initially reviewed. Slides that incorporated the invasive margin of the tumour were selected. Sections were then deparaffinised and hydrated. After antigen retrieval, the sections were incubated with CD8 (C8/144B; Dako; 1:100 dilution) and CD3 (2GV6, Ventana) antibodies. ImmunoCruz mouse ABC staining system (Santa Cruz Biotechnology) for CD8 and ultraView Universal DAB detection kit (Ventana) for CD3 were used for secondary antibody. Slides were digitally conserved using the Aperio. Three non-contiguous areas of highest lymphocyte density were selected at both the CT and IM. The number of CD3+ and CD8+ TILs in each selected area (CD3CT, CD3IM, CD8CT, CD8IM) were estimated using Aperio image analysis software. The average of three non-contiguous areas was used to eliminate sampling error[11 (link), 13 (link)]. The reviewer who selected the areas for analysis was blinded to the clinical outcome of the patient.
Immunocruz mouse abc staining system
Manufactured by Santa Cruz Biotechnology
Sourced in China
The ImmunoCruz mouse ABC staining system is a laboratory equipment used for the detection and visualization of target antigens in mouse tissue samples. It utilizes an avidin-biotin complex (ABC) method to amplify the signal, providing enhanced sensitivity. The system includes pre-optimized reagents and a step-by-step protocol to facilitate the staining process.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using immunocruz mouse abc staining system
1
Quantifying CD3+ and CD8+ TILs in Bladder Cancer
2
Immunohistochemical Analysis of IRGM in Melanoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embedded sections were subjected to antigen retrieval with citrate buffer (pH 6.0), incubated with mouse anti-human IRGM monoclonal antibody (Ab; 1G9; AbMart, Shanghai, China; 1:200), and visualized using the ImmunoCruz mouse ABC staining system (sc-2017; Santa Cruz Biotechnology, Shanghai, China). Evaluation of immunohistochemical scores was not performed for cases with severe melanin diffusion in human melanoma tissue chips. In the 87 melanoma cases obtained from Harbin Medical University Cancer Hospital, diffused melanin was initially removed with oxalic acid and potassium permanganate before IHC staining.
In terms of the immunohistochemical score standard, a brown signal was indicative of positive action. The stain index was determined as a product of intensity and proportion, ranging from 0 to 12. Intensity scores were classified as 0 (negative), 1 (weak), 2 (moderate), or 3 (strong). The proportion, representing the frequency of positive cells, was scored as 0 (<5%), 1 (5%–25%), 2 (26%–50%), 3 (51%–75%), and 4 (>75%). We randomly selected three points in each case and calculated the stain index of each point. The stain index in each case was determined as the average of three points. For statistical analyses, scores of 0–6 were considered as “low expression” and 7–12 as “high expression.”
In terms of the immunohistochemical score standard, a brown signal was indicative of positive action. The stain index was determined as a product of intensity and proportion, ranging from 0 to 12. Intensity scores were classified as 0 (negative), 1 (weak), 2 (moderate), or 3 (strong). The proportion, representing the frequency of positive cells, was scored as 0 (<5%), 1 (5%–25%), 2 (26%–50%), 3 (51%–75%), and 4 (>75%). We randomly selected three points in each case and calculated the stain index of each point. The stain index in each case was determined as the average of three points. For statistical analyses, scores of 0–6 were considered as “low expression” and 7–12 as “high expression.”
3
Monitoring Neural Stemness Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were fixed at different periods (after 24, 25, 27, and 29 hours) for the immunocytochemistry assay for each NES, NF-L, and MAP-2 markers. This helped to detect neural stemness state from the protein level. The cells were fixed and stained using the Immuno-Cruz mouse ABC Staining System (Santa Cruz Biotechnology Inc.) at all times. Staining was done using specific antibodies with NES as a neural stemness marker, NF-L as an early neural cells marker, and MAP-2 as a mature neural marker. The immunocytochemistry assay was carried out according to the manufacturer’s instructions.
4
Immunohistochemical Analysis of CD20 in Plasmacytoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The plasmacytoma cells were cultured on coverslips. The cells were allowed to develop a monolayer. Then, the cells were fixed with 4% neutral buffer formalin for 10 minutes, washed with phosphate-buffered saline (PBS), and left to dry. After fixation, slides were incubated in a humidified chamber with 1% H2O2 for 10 minutes, washed two to three times with PBS, and incubated with 1.5% blocking serum for 30–40 minutes at room temperature. Then, the primary antibody of the rabbit anti-CD20 (Santa Cruz Biotechnology Inc., Dallas, TX, USA) was incubated overnight at 4°C in a humidified chamber. Afterward, it was stained by the ImmunoCruz™ mouse ABC staining system (Santa Cruz Biotechnology). The biotinylated secondary antibody was incubated for 30 minutes, and then washed. Avidin horseradish peroxidase was then added and incubated for another 30 minutes, then washed. Liquid 3,3′-diaminobenzidine chromogen-solution formula was added to each slide for 20 minutes at room temperature, washed extensively with PBS, and counterstained with hematoxylin for 30–60 seconds. The slides were mounted with Distyrene, a plasticizer, and xylene (DPX) (supplied by Sigma-Aldrich Co., St Louis, MO, USA), inspected using light microscopy, and photographed using a digital camera.
5
Immunohistochemical Analysis of Liver Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver tissue was formaldehyde-fixed and paraffin-embedded in blocks. 5 µm thick liver sections were de-paraffined, rehydrated, then heated to 100ºC for 5 min in citrate buffer (pH 6.0) for antigen retrieval. Immunohistochemistry for proliferating cell nuclear antigen (PCNA; Santa Cruz, sc-25280) was performed using the ImmunoCruz Mouse ABC Staining System (Santa Cruz) according to the manufacturer's instructions. Liver sections were stained for haematoxylin/eosin (HE) and steatosis was double-blind quantified from low (0) to high (5) steatosis.
For Ki67 immunostaining, liver tissue was formaldehyde-fixed for 24 h and 30% sucrose cryopreserved for 24 h. The sample was embedded in OCT (Tissue-Tek) at -80ºC. Liver sections (10 µm thick) were blocked with 1% bovine serum albumin for 20 min and incubated with rabbit anti-Ki67 (Abcam, ab15580) and secondary antibody goat anti-rabbit-IgG-A647 (Life Technologies, A21244). The samples were mounted in Mowiol (Calbiochem).
For Ki67 immunostaining, liver tissue was formaldehyde-fixed for 24 h and 30% sucrose cryopreserved for 24 h. The sample was embedded in OCT (Tissue-Tek) at -80ºC. Liver sections (10 µm thick) were blocked with 1% bovine serum albumin for 20 min and incubated with rabbit anti-Ki67 (Abcam, ab15580) and secondary antibody goat anti-rabbit-IgG-A647 (Life Technologies, A21244). The samples were mounted in Mowiol (Calbiochem).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!