Regional multidisciplinary team intervention programme to improve colorectal cancer outcomes: study protocol for the Yorkshire Cancer Research Bowel Cancer Improvement Programme (YCR BCIP).

Although colorectal cancer outcomes in England are improving, they remain poorer than many comparable countries. Yorkshire Cancer Research has, therefore, established a Bowel Cancer Improvement Programme (YCR BCIP) to improve colorectal cancer outcomes within Yorkshire and Humber, a region representative of the nation. It aims to do this by quantifying variation in practice, engaging with the colorectal multidisciplinary teams (MDTs) to understand this and developing educational interventions to minimise it and improve outcomes.Initially, routine health datasets will be used to quantify variation in the demographics, management and outcomes of patients across the Yorkshire and Humber region and results presented to MDTs. The YCR BCIP is seeking to supplement these existing data with patient-reported health-related quality of life information (patient-reported outcome measures, PROMs) and tissue sample analysis.
Specialty groups (surgery, radiology, pathology, clinical oncology, medical oncology, clinical nurse specialists and anaesthetics) have been established to provide oversight and direction for their clinical area within the programme, to review data and analysis and to develop appropriate educational initiatives.The YCR BCIP is aiming to address the variation in practice to significantly improve colorectal cancer outcomes across the Yorkshire and Humber region. PROMs and tissue sample collection and analysis will help to capture the information required to fully assess care in the region. Engagement of the region’s MDTs with their data will lead to a range of educational initiatives, studies and clinical audits that aim to optimise practice across the region.

SEAP activity measurement in reporter cell-based assays using BCIP / NBT as substrate.

SEAP (secreted embryonic alkaline phosphatase) has been suggested as versatile reporter protein inter alia for cell ligand interaction. Generic photometric assay formats for this enzyme are currently lacking. Using the interaction of recombinant hCD40 ligand with HEK-Blue sensor cells expressing the CD40 receptor as example, we show that such an assay can be developed based on BCIP/NBT (5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium chloride) as substrate. Supplementation of the reaction buffer with a micelle-forming detergent (TWEEN 20) stabilizes the water-insoluble reactions products thereby allowing reproducible photometric quantification of the colloidal dispersion.
After optimizing the assay in terms of incubation time, cell number and environmental conditions, a cellular response to stimulation was already visible for 0.25 ng mL-1 of rhCD40L. Moreover, the sensitivity of the assay was significantly better than reported previously for alternative assays used in combination with the commercially available reporter cells. The use of BCIP/NBT as substrate therefore provides a robust and sensitive method to monitor SEAP activity in solution, which could conceivably be extended to other cell-based and biological assays using SEAP as reporter protein.

BCIP: a gene-centered platform for identifying potential regulatory genes in breast cancer.

Breast cancer is a disease with high heterogeneity. Many issues on tumorigenesis and progression are still elusive. It is critical to identify genes that play important roles in the progression of tumors, especially for tumors with poor prognosis such as basal-like breast cancer and tumors in very young women. To facilitate the identification of potential regulatory or driver genes, we present the Breast Cancer Integrative Platform (BCIP, http://omics.bmi.ac.cn/bcancer/). BCIP maintains multi-omics data selected with strict quality control and processed with uniform normalization methods, including gene expression profiles from 9,005 tumor and 376 normal tissue samples, copy number variation information from 3,035 tumor samples, microRNA-target interactions, co-expressed genes, KEGG pathways, and mammary tissue-specific gene functional networks.
This platform provides a user-friendly interface integrating comprehensive and flexible analysis tools on differential gene expression, copy number variation, and survival analysis. The prominent characteristic of BCIP is that users can perform analysis by customizing subgroups with single or combined clinical features, including subtypes, histological grades, pathologic stages, metastasis status, lymph node status, ER/PR/HER2 status, TP53 mutation status, menopause status, age, tumor size, therapy responses, and prognosis. BCIP will help to identify regulatory or driver genes and candidate biomarkers for further research in breast cancer.

Cellular resolution expression profiling using confocal detection of NBT/BCIP precipitate by reflection microscopy.

The determination of gene expression patterns in three dimensions with cellular resolution is an important goal in developmental biology. However the most sensitive, efficient, and widely used staining technique for whole-mount in situ hybridization (WMISH), nitroblue tetrazolium (NBT)/5-bromo-4-chloro-3-indolyl phosphate (BCIP) precipitation by alkaline phosphatase, could not yet be combined with the most precise, high-resolution detection technique, confocal laser-scanning microscopy (CLSM).
Here we report the efficient visualization of the NBT/BCIP precipitate using confocal reflection microscopy for WMISH samples of Drosophila, zebrafish, and the marine annelid worm, Platynereis dumerilii. In our simple WMISH protocol for reflection CLSM, NBT/BCIP staining can be combined with fluorescent WMISH, immunostainings, or transgenic green fluorescent protein (GFP) marker lines, allowing double labeling of cell types or of embryological structures of interest. Whole-mount reflection CLSM will thus greatly facilitate large-scale cellular resolution expression profiling in vertebrate and invertebrate model organisms.

Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.

In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP).
The solid reaction product has significant fluorescence that enables the use of confocal microscopy to generate high-resolution three-dimensional (3-D) imaging of gene expression.
A modified Ce/Mg-BCIP-NBT formazan/indigoblue technique for demonstration of non-specific alkaline phosphatase activity.

The wide ranged structurally variability of formazans and their accessibility for auxiliary additives as redoxmediators or metals provide an easy tunable chromogenic visualization technique. We present here an improved nitro blue tetrazolium (NBT) 5-bromo-4-chloro-3-indolyl phosphate (BCIP) method which is superior to the classical McGadey’s procedure regarding proper precipitation and localization as well as sensitivity. Different metal additives as well as the overall reaction course modifying additives (redox mediators, chelating additives, buffer) were optimized.

BCIP

HY-15909 MedChemExpress 500mg 271 EUR

BCIP

K2191050-4 Biochain 150 ul 137 EUR

BCIP/NBT

CH015 ABM 100 ml 131 EUR

BCIP/NBT Substrate

F064-100 Cygnus Technologies 100 ml 257 EUR

BCIP/NBT Substrate

F064-1000 Cygnus Technologies 1000 ml 811 EUR

BCIP toluidine salt

BB0072 Bio Basic 500mg 134.39 EUR

BCIP disodium salt

BB1171 Bio Basic 500mg 232.7 EUR

BCIP/NBT kit

10003 Biotium 1SET 185 EUR

BCIP/INT Solution

ACL050 ScyTek Laboratories 50 ml 79 EUR

BCIP/INT Solution

ACL125 ScyTek Laboratories 125 ml 98 EUR

BCIP/INT Solution

ACL500 ScyTek Laboratories 500 ml 205 EUR

BCIP/INT Solution

ACL999 ScyTek Laboratories 1000 ml 319 EUR

BCIP/NBT Solution

ACN050 ScyTek Laboratories 50 ml 79 EUR

BCIP/NBT Solution

ACN125 ScyTek Laboratories 125 ml 117 EUR

BCIP/NBT Solution

ACN500 ScyTek Laboratories 500 ml 260 EUR

BCIP/NBT Solution

ACN999 ScyTek Laboratories 1000 ml 394 EUR

BCIP/NBT Substrate

42-BC07 Fitzgerald 100 ml 129 EUR

BCIP (Molecular Biology Grade)

CE108 GeneOn 250 mg 63 EUR

BCIP (Molecular Biology Grade)

CE109 GeneOn 1 g 90 EUR

BCIP p-Toluidine Salt

abx082537-100mg Abbexa 100 mg 203 EUR

NBT/BCIP Stain Kit

PW032 Bio Basic 5Preps, 5prep 70.88 EUR

INT/BCIP Stain Kit

PW033 Bio Basic 5Preps, 5prep 70.88 EUR

BCIP red/NBT kit

10005 Biotium 1SET 219 EUR

BCIP pink/NBT kit

10007 Biotium 1SET 239 EUR

BCIP/ NBT Chromogenic Substrate Kit

AR1023 BosterBio 1 kit (20X) 103 EUR

BCIP/NBT Solution for IHC

B3007-005 GenDepot 50ml 134 EUR

BCIP/NBT Solution for IHC

B3007-010 GenDepot 100ml 188 EUR

BCIP/NBT Solution for IHC

B3007-050 GenDepot 500ml 475 EUR

BCIP-NBT Dye for IHC

G447 ABM 250 ml 254 EUR

BCIP, 5-Bromo-4-Chloro-3-Indolyl Phosphate

CH012 ABM 100 mg 128 EUR

U-3′-BCIP: a chromogenic substrate for the detection of RNase A in recombinant DNA expression systems.

The synthesis of the bovine pancreatic ribonuclease A (RNase A, EC 3.1.27.5) chromogenic substrate uridine-3′-(5-bromo-4-chloroindol-3-yl)-phosphate (U-3′-BCIP) is described. RNase A catalyzes the hydrolysis of U-3′-BCIP to release a halogenated indol-3-ol that undergoes rapid aerobic oxidation to the dark blue 5,5′-dibromo-4,4′-dichloroindigo. Preliminary kinetic studies indicate that this compound may have practical use for assaying RNase A activity both in vitro and in vivo, e.g. in screening bacterial colonies for RNase A produced by recombinant DNA methods.

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