Human breast tumor tissue array (BC081116d) with known ERα status was obtained from US Biomax (Derwood, MD). The sections were deparaffinized and rehydrated through graded ethanol. Antigen retrieval was performed using citrate buffer pH 6.0 prior to incubation with 3% H2O2 to reduce endogenous peroxidase activity and blocking with goat serum. The slides were then incubated with anti-AKTIP or anti-ERα antibody overnight at 4°C followed by biotin-conjugated secondary antibody (Dako, Carpinteria, CA) incubation at room temperature for 1 hr. 3,3′-diaminobenzidine (DAB, Amresco, Solon, OH) was used to detect signal from HRP. Histoscores on an arbitrary scale: 0, no immunoreactivity; 1, weak; 2, moderate; 3, intense; and 4, very intense were used to represent protein expression of AKTIP whereas histoscores of 0, no immunoreactivity; 1, weak; 2, moderate; 3, intense were used to represent protein expression of ERα. Our ERα staining scores are completely concordant with the reported ERα intensity provided by the company.
Bc081116d
Manufactured by Tissue Array
Sourced in United States
BC081116d is a laboratory equipment product. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
Automatically generated - may contain errors
4 protocols using bc081116d
1
Immunohistochemical Analysis of AKTIP and ERα
2
Immunohistochemical Analysis of AKTIP and ERα
3
Immunohistochemical Analysis of Breast Carcinoma
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The formalin-fixed paraffin-embedded human breast carcinoma tissue microarray containing 100 cases of invasive carcinoma of no special type was obtained from US Biomax (BC081116d, US Biomax Inc., Rockville, MD, USA). Before deparaffinization with xylene, slides were baked in an incubator at 60 °C for 1 h. Rehydration was performed with gradually reduced concentrations of ethanol from 100% to 95%, 85%, and 70% for 1 m. For antigen retrieval, the slides were submerged in citrate buffer and boiled in a microwave for 10 m. Sufficiently cooled slides were stained using Novolink Polymer Detection Systems (Leica Biosystems Inc., Buffalo Grove, IL, USA) according to the manufacturer's instructions. The anti-HOXB2 (1:200, ab220390, Abcam), anti-MATN3 (1:100, ab238893, Abcam), and anti-ECM2 (1:300, ab122268, Abcam) antibodies were diluted with phosphate buffered saline with Tween 20 (PBST) buffer. DAB staining was carried out until the tissue sections expressed a brown color. Tissue samples were then dehydrated with 70%, 85%, 95%, and 100% ethanol and cleared with xylene. The DAB signals of 100 breast carcinomas were scored using ImageJ software. Negative/low and high levels were classified based on the score of each protein signal intensity and area.
4
STARD3 Immunohistochemistry in TMAs
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We used two tissue microarrays (TMA) provided from, US-Biomax; BC081116d and BC081120f. Primary antibody against STARD3 was purchased from Abcam [anti-MLN64 antibody (ab3478)]. The Mouse and Rabbit Specific HRP/DAB immunohistochemistry (IHC) Detection Kit was purchased from Abcam (cat number: ab64261).
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