This document summarizes a study evaluating the efficacy of differentiating mouse embryonic stem cells (mESCs) into hepatocyte-like cells. The researchers found that cell cycle synchronization of mESCs prior to differentiation led to decreased expression of the pluripotency marker Oct4 and increased expression of endoderm and hepatic progenitor markers, indicating improved differentiation. Assays of stem cell-derived hepatocyte-like cells showed production of the liver proteins albumin and urea, though urea levels decreased over time, suggesting further optimization is needed to generate fully functional hepatocytes from mESCs. The researchers concluded cell cycle synchronization enhanced differentiation but that identifying additional progenitor factors could improve hepatic function of the derived cells.
Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different subpopulations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell.
Fa cs sample preparation & protocolJames Waita
Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different subpopulations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell.
Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. Mammalian cell culture technology has become a major field in modern biotechnology; mammalian cell culture refers to the cells of a mammalian, isolated from specific tissues (i.e. skin, liver, glands, etc.) and further cultivated and reproduced in an artificial medium. Cell culture technology is currently playing a major role in toxicity testing, cancer research, virology, genetic engineering, and gene therapy.
OBJECTIVE:
To observe the transfection of CHO and HEK cells with GFP
To observe the recombinant GFP using Western Blotting
To purify the transfected HEK and CHO cells using AKTA Pure Purification
The occurrence of AmpC β-lactamase and ESBL producing Gram-negative bacteria ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different subpopulations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell.
Fa cs sample preparation & protocolJames Waita
Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different subpopulations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell.
Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. Mammalian cell culture technology has become a major field in modern biotechnology; mammalian cell culture refers to the cells of a mammalian, isolated from specific tissues (i.e. skin, liver, glands, etc.) and further cultivated and reproduced in an artificial medium. Cell culture technology is currently playing a major role in toxicity testing, cancer research, virology, genetic engineering, and gene therapy.
OBJECTIVE:
To observe the transfection of CHO and HEK cells with GFP
To observe the recombinant GFP using Western Blotting
To purify the transfected HEK and CHO cells using AKTA Pure Purification
The occurrence of AmpC β-lactamase and ESBL producing Gram-negative bacteria ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Keratin 16 is a protein that in humans is encoded by the KRT16 gene.
Keratin 16 is a type I cytokeratin. It is paired with keratin 6 in a number of epithelial tissues, including nail bed, esophagus, tongue, and hair follicles. Mutations in the gene encoding this protein are associated with the genetic skin disorders pachyonychia congenita, non-epidermolytic palmoplantar keratoderma and unilateral palmoplantar verrucous nevus.
Anti-CK16 -http://www.stjohnslabs.com/ck16-antibody-p-98593
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Keratin-7 is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues. This type II cytokeratin is specifically expressed in the simple epithelia lining the cavities of the internal organs and in the gland ducts and blood vessels. The genes encoding the type II cytokeratins are clustered in a region of chromosome 12q12-q13. Alternative splicing may result in several transcript variants; however, not all variants have been fully described.
Anti-CK7 -http://www.stjohnslabs.com/ck7-antibody-p-98591
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Production of recombinant antibodies by hostEchoHan4
Digoxin Immune Fab is a sheep antibody (26-10) FAB fragment from sheep immunized with the digoxin derivative Digoxindicarboxymethylamine. It is used as an antidote for overdose of digoxin.https://www.creativebiolabs.net/Digoxin-Immune-Fab-Ovine-22441.htm
Immunofluorescence Antibody Validation Report for Anti-CA IX Antibody (STJ96978)St John's Laboratory Ltd
Reversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia.
Anti-CA IX -http://www.stjohnslabs.com/ca-ix-antibody-p-98613
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP). Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9. Strict requirement for an Asp residue at position P1 and with a marked preference for His at position P2. It has a preferred cleavage sequence of Leu-Gly-His-Asp-|-Xaa. Inhibited by the effector protein NleF that is produced by pathogenic E.coli; this inhibits apoptosis.
Anti-Caspase-9 -http://www.stjohnslabs.com/caspase-9-antibody-p-91489
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Immunofluorescence Antibody Validation Report for Anti-Collagen IV Antibody (...St John's Laboratory Ltd
Type IV collagen is the major structural component of glomerular basement membranes (GBM), forming a 'chicken-wire' meshwork together with laminins, proteoglycans and entactin/nidogen.; Arresten, comprising the C-terminal NC1 domain, inhibits angiogenesis and tumor formation. The C-terminal half is found to possess the anti-angiogenic activity. Specifically inhibits endothelial cell proliferation, migration and tube formation. Inhibits expression of hypoxia-inducible factor 1alpha and ERK1/2 and p38 MAPK activation. Ligand for alpha1/beta1 integrin.
Anti-Collagen IV - http://www.stjohnslabs.com/anti-collagen-iv-antibody?filter_name=STJ98907
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Indirect flow cytometry, also known as fluorescence-activated cell sorting (FACS) is a specialised type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell. It is a useful scientific instrument as it provides fast, objective and quantitative recording of fluorescent signals from individual cells as well as physical separation of cells of particular interest.
In contrast to direct flow cytometry, FACS requires two incubation steps. Firstly with the appropriate primary antibody, then with a fluorochrome-labelled secondary antibody.
Keratin 16 is a protein that in humans is encoded by the KRT16 gene.
Keratin 16 is a type I cytokeratin. It is paired with keratin 6 in a number of epithelial tissues, including nail bed, esophagus, tongue, and hair follicles. Mutations in the gene encoding this protein are associated with the genetic skin disorders pachyonychia congenita, non-epidermolytic palmoplantar keratoderma and unilateral palmoplantar verrucous nevus.
Anti-CK16 -http://www.stjohnslabs.com/ck16-antibody-p-98593
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Keratin-7 is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues. This type II cytokeratin is specifically expressed in the simple epithelia lining the cavities of the internal organs and in the gland ducts and blood vessels. The genes encoding the type II cytokeratins are clustered in a region of chromosome 12q12-q13. Alternative splicing may result in several transcript variants; however, not all variants have been fully described.
Anti-CK7 -http://www.stjohnslabs.com/ck7-antibody-p-98591
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Production of recombinant antibodies by hostEchoHan4
Digoxin Immune Fab is a sheep antibody (26-10) FAB fragment from sheep immunized with the digoxin derivative Digoxindicarboxymethylamine. It is used as an antidote for overdose of digoxin.https://www.creativebiolabs.net/Digoxin-Immune-Fab-Ovine-22441.htm
Immunofluorescence Antibody Validation Report for Anti-CA IX Antibody (STJ96978)St John's Laboratory Ltd
Reversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia.
Anti-CA IX -http://www.stjohnslabs.com/ca-ix-antibody-p-98613
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP). Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9. Strict requirement for an Asp residue at position P1 and with a marked preference for His at position P2. It has a preferred cleavage sequence of Leu-Gly-His-Asp-|-Xaa. Inhibited by the effector protein NleF that is produced by pathogenic E.coli; this inhibits apoptosis.
Anti-Caspase-9 -http://www.stjohnslabs.com/caspase-9-antibody-p-91489
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Immunofluorescence Antibody Validation Report for Anti-Collagen IV Antibody (...St John's Laboratory Ltd
Type IV collagen is the major structural component of glomerular basement membranes (GBM), forming a 'chicken-wire' meshwork together with laminins, proteoglycans and entactin/nidogen.; Arresten, comprising the C-terminal NC1 domain, inhibits angiogenesis and tumor formation. The C-terminal half is found to possess the anti-angiogenic activity. Specifically inhibits endothelial cell proliferation, migration and tube formation. Inhibits expression of hypoxia-inducible factor 1alpha and ERK1/2 and p38 MAPK activation. Ligand for alpha1/beta1 integrin.
Anti-Collagen IV - http://www.stjohnslabs.com/anti-collagen-iv-antibody?filter_name=STJ98907
Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
Indirect flow cytometry, also known as fluorescence-activated cell sorting (FACS) is a specialised type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell. It is a useful scientific instrument as it provides fast, objective and quantitative recording of fluorescent signals from individual cells as well as physical separation of cells of particular interest.
In contrast to direct flow cytometry, FACS requires two incubation steps. Firstly with the appropriate primary antibody, then with a fluorochrome-labelled secondary antibody.
German Scientist “Carl Vogt” was first to describe the principle of apoptosis in 1842. In 1885, Anatomist “Walther Flemming” gave more precise description of Programmed Cell Death. Apoptosis is a form of Programmed Cell Death that occurs in multicellular organisms. It is a Greek word which means falling off. It leads to breakdown and disposal of cells. Macrophages and other Phagocytic Cells remove them by Phagocytosis, without developing any type of inflammation. It is a biochemical event that leads to morphological changes and death. The average adult human looses 50-70 billion cells each day due to apoptosis.
Blood grouping of liquid blood: forward typing and reverse typing; Blood grouping of dried blood: Lattes test, adsorption-elution, adsorption-inhibition, mixed agglutination; HLA antigens and HLA typing; Role of sero-genetic markers in individualization and paternity disputes; Pitfalls in red cell typing
Onion (Allium Cepa) Genotoxicity Test
Laboratory of Ecotoxicology and LCA
Department of Environmental Chemistry, ICT Prague
References:
1. FERETTI, D., ZERBINI, I., ZANI, C., CERETTI, E., MORETTI,M.,MONARCA, S. (2007): Allium cepa chromosome
abberation and micronucleus tests applied to study genotoxicity of extracts from pesticide-treated vegetables and
grapes. Food Addit. Contam. 24 (6): 561-572.
2. RANK, J., NIELSEN, M.H. (1997): Allium anaphase-telophase genotoxicity assay. Department of Environment,
Technology and Social Studies, Roskilde University, Denmark.
1 Objectives Genetically transform bacteria with for.docxmercysuttle
1
Objectives
Genetically transform bacteria with
foreign DNA and induce
expression of genes encoded on
DNA to produce novel
Isolate chromosomal DNA from
Introduction
In this portion of the lab, you will perform a
procedure known as genetic
transformation. that a gene is
a piece of DNA that provides the
instructions for making (codes for) a
protein. This gives an organism a
particular trait. Genetic transformation
literally means change caused by genes,
involves the insertion of a gene into an
organism in order to change the organism’s
trait. transformation is used in
many areas of biotechnology. In
agriculture, genes coding for traits such as
pest, or spoilage resistance can be
genetically transformed into plants. In
bioremediation, bacteria can genetically
transformed with genes enabling them to
digest oil spills. In medicine, diseases
caused defective genes are beginning
to be treated by gene therapy; that is, by
genetically transforming a person’s
cells with healthy copies of the defective
gene that causes the
You will use a procedure to transform
bacteria with a gene that codes for Green
Fluorescent (GFP). The real-life
source of this gene is the bioluminescent
jellyfish Aequorea victoria.
Fluorescent Protein causes the jellyfish to
fluoresce and glow in the dark.
LAB TOPIC 10: Nucleic Acids and Genetic Transformation
Following the procedure,
the bacteria express their newly acquired
jellyfish gene and produce the fluorescent
which causes them to glow a
brilliant green color under ultraviolet
In this activity, you will learn about the
process of moving genes from one organism
to another with aid of a plasmid. In
nature, bacteria can transfer plasmids back
and forth allowing them to share
beneficial genes. This natural mechanism
allows bacteria to adapt to new
environments. The occurrence of
bacterial resistance to is due to
the transmission of
Genetic transformation involves
insertion of some new DNA into the E.
cells. In addition to one large
bacteria often contain one or more
circular pieces of DNA called
Plasmid DNA usually contains genes for
than one trait. Scientists can use a
called genetic engineering to insert
coding for new traits into a plasmid.
In case, the pGLO plasmid carries the
GFP that codes for the green
fluorescent protein and a gene (bla) that
codes for a protein that gives the
resistance to an antibiotic. The genetically
engineered plasmid can then be used to
genetically bacteria to give them
this new
Figure 10.1 Bacterial cell undergoing genetic transformation with the pGLO
plasmid
Exercise 10.1
Bacterial Transformation
2
Pre-lab exercises:
Since scientific laboratory investigations
are designed to get information about a
question, our first might be to
formulate some questions for this
Can we genetically transform an organism?
Which organism is
1. To genetically tra ...