ABSTRACT
Antibodies are the most widely used reagent for isolation and detection of specific proteins. However, using antibodies that are not highly specific in these studies can generate inaccurate and misleading data. Protein microarrays offer a platform by which antibody cross-reactivity against a broad range of cellular antigens can be simultaneously and quantitatively profiled. This protocol describes in detail the process of array pretreatment, antibody binding, washing, scanning and quantitative analysis of antibody specificity.
Subject(s)
Antibody Specificity/immunology , Protein Array Analysis/methods , Proteomics/methods , Animals , Antibodies/genetics , Antibodies/immunology , Antigens/genetics , Antigens/immunology , HumansABSTRACT
A key component of efforts to address the reproducibility crisis in biomedical research is the development of rigorously validated and renewable protein-affinity reagents. As part of the US National Institutes of Health (NIH) Protein Capture Reagents Program (PCRP), we have generated a collection of 1,406 highly validated immunoprecipitation- and/or immunoblotting-grade mouse monoclonal antibodies (mAbs) to 737 human transcription factors, using an integrated production and validation pipeline. We used HuProt human protein microarrays as a primary validation tool to identify mAbs with high specificity for their cognate targets. We further validated PCRP mAbs by means of multiple experimental applications, including immunoprecipitation, immunoblotting, chromatin immunoprecipitation followed by sequencing (ChIP-seq), and immunohistochemistry. We also conducted a meta-analysis that identified critical variables that contribute to the generation of high-quality mAbs. All validation data, protocols, and links to PCRP mAb suppliers are available at http://proteincapture.org.
Subject(s)
Antibodies, Monoclonal/immunology , Protein Array Analysis/methods , Transcription Factors/metabolism , Animals , Cloning, Molecular , Databases, Factual , Female , HeLa Cells , Humans , Mice , Mice, Inbred BALB C , Reproducibility of ResultsABSTRACT
BACKGROUND: The transition to practice for new graduate nurses poses distinct challenges at the point of care. The complicated work environment necessitates a preceptorship model that maintains safety at all times. METHOD: This study used survey, focus groups, and interviews to assess the effects of the married state preceptorship model (MSPM) on the experiences and perceptions of the new nurses and preceptors in a hospital setting. RESULTS: The result validated the original findings from the new nurses' survey, indicating all (N = 28, 100%) of the new nurses found the MSPM to be beneficial, promote safety, and lessen anxiety, and 27 (96%) thought it helped them to assume a full patient load. The preceptors' survey revealed that 29 (97%) of the preceptors thought the MSPM promoted safety, 30 (100%) thought it boosted confidence, and 30 (100%) believed it meet the needs of the new nurses. CONCLUSION: The study showed same thematic saturation on partnership, critical thinking, learning, and transition. The change in culture has been strengthened in the replicate study referring to their acclimatization in the unit through the MSPM. J Contin Educ Nurs. 2016;47(11):511-517.
Subject(s)
Attitude of Health Personnel , Clinical Competence , Inservice Training/organization & administration , Nursing Staff, Hospital/psychology , Preceptorship/organization & administration , Adult , Female , Focus Groups , Hospitals, Community , Humans , Male , Middle Aged , Models, Nursing , Northwestern United States , Surveys and QuestionnairesABSTRACT
A 68-year-old athletic woman presented to our institution in January 2002 with a several-month history of progressing complaints of pain, swelling, and loss of motion in the right knee. These manifestations had begun the previous July during a game of tennis. She experienced persisting pain and recurring effusions. Because the patient had been residing in another state between July and January, rheumatologic and orthopaedic evaluations of the knee, including a magnetic resonance imaging (MRI), had been performed at a geographically distant (but affiliated) institution. The resulting presumptive diagnosis was a "wear and tear" degenerative articular disorder of the knee. A program of anti-inflammatory medication and physical therapy was begun for several months but produced no therapeutic benefit by the time the patient presented at our institution. After examination confirmed marked losses of both flexion and extension of the knee, effusion, and exquisite medial joint tenderness, an MRI was repeated, using intra-articular gadolinium as a contrast agent. It revealed an intra-articular mass encircling the medial and posterior extents of the medial femoral condyle. An arthroscopic multiportal excisional biopsy was performed. It revealed the existence of a juxta-articular myxoma. The patient recovered most of the range of motion during the next several months, and the effusion and severe pain gradually dissipated. The patient was subsequently followed by sequential physical examinations and MRIs, performed at increasing intervals of time, without recurrence of a mass or of her flagrant symptoms in the first year post surgery. Though the patient's diagnosis was established and treatment outcome was satisfactory, many issues were brought up in this case regarding most appropriate selection of diagnostic tests and treatment approaches.
