Wednesday, 24 June 2015

Platelet function assessment in clinical trials

Do you need to consider platelet aggregation? 

It is my pleasure to introduce Java Clinical Research. A Clinical Research Organisation that has supported the assessment of platelet function in clinical trials since 1999.

Platelet function studies provide vital pharmacodynamic and population response data. This is often required by regulatory agencies if:

  • Your drug directly affects platelet function (e.g. ADP antagonists).
  • Your drug is in a class that has been shown to affect platelet function as a side effect (e.g. 5-HT4 antagonists).
  • Your drug's target suggests it may alter platelet function (e.g. BTK inhibitors).

Java Clinical offers a complete service in the design, implementation and interpretation of platelet function assays.

How can Java Clinical help?

Java Clinical offers guidance at each stage in the planning, execution and analysis of platelet assay services in preclinical and clinical trials. Full clinical trial management is also provided, with strategic advice on the clinical trial approval process, project management, regulatory application services, and clinical trial monitoring for all types of clinical studies. For more detail on how Java Clinical can help, please click on the links below:

Wednesday, 17 June 2015

TransFix - A Cellular Antigen Stabilisation Reagent

What is TransFix?

TransFix is a stabilisation solution that stabilises cellular suspensions of both human and animal origin whilst preserving the integrity and antigenicity of leukocytes for transportation and storage. The components of TransFix stabilise cellular markers, allowing analysis by flow cytometry to be carried out at a more convenient time for the user. TransFix can be used in clinical diagnostic testing, clinical trials and a wide range of research applications…

Human Blood Stabilisation
For immunophenotyping, untreated blood samples have a viable shelf life of 24-48 hours due to enzymatic degradation of cellular markers. Treating blood samples with TransFix enables flow cytometric analysis of extracellular markers at 2°C to 25°C for 10 days and at 37°C for 3 days.

TransFix is available in several formats including TransFix Sample Storage Tubes and TransFix/EDTA Vacuum Blood Collection Tubes (direct draw collection tubes used to stabilise venous blood at the point of collection and preserve whole blood specimens for immunophenotyping by flow cytometry). TransFix is also available in bulk volumes.

Read More...

CSF Stabilisation
TransFix/EDTA CSF Sample Storage Tubes have been developed in collaboration with research oncologists and contain TransFix that has been optimised for CSF leukocyte stabilisation.

Ordinarily, CSF samples require flow cytometric analysis within 2 hours of lumbar puncture, as the immunophenotypic profile is detrimentally affected by natural degradation. The components of the TransFix reagent used in TransFix/EDTA CSF Sample Storage Tubes stabilise CSF leukocyte markers, making it possible to determine cell subsets by flow cytometric analysis up to 10 days after collection.

A recent paper in the British Journal of Haematology (Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms, Johansson et al, 2014) recommended that unless immediate analysis is possible then CSF samples can be stored in TransFix/EDTA CSF Sample Storage Tubes for up to 72 hours before analysis.

Read More...

 Bone Marrow Stabilisation
TransFix has been shown to stabilise cells within human bone marrow populations. This allows immunophenotyping by flow cytometry to be conducted at a time that is more convenient to the operator.

Independent research involving the use of TransFix to stabilise bone marrow samples has been published, including the use of TransFix for the enumeration of bone marrow mast cells  and for immunophenotyping rat bone marrow stem cells

Read More…

Immunostaining of Circulating Tumour Cells
The presence of circulating tumour cells (CTC’s) has been shown to be of diagnostic relevance for patients with early and advanced breast cancer. The usefulness of CTC assessments depends upon accurate cell counts and the corresponding analysis of molecular targets.
TransFix stabilised blood samples were used for CTC analysis in an independent study. CTCs were detected by image analytics after fluorescence scanning microscopy.

Read More...

Stabilisation of Animal Blood
Researchers have found that the addition of TransFix to animal blood samples at the time of collection has significantly extended the integrity of the samples in several different species.
Independent research studies involving the use of TransFix in stabilising animal blood samples have been published:
- A rapid high precision flow cytometry based technique for total white blood cell counting in chickens
- Effects of Cyclosporin A induced T-lymphocyte depletion on the course of avian Metapneumovirus (aMPV) infection in turkeys. 
TransFix has also been used to successfully stabilise whole blood from the following species: mouse, rat, guinea pig, sheep, seal, turtle, ostrich and pig. However, these studies have yet to be published.

Read More…

TransFix is produced by Cytomark - a division of Caltag Medsystems. If you would like further information about TransFix, please do not hesitate to contact us at cytomark@caltagmedsystems.co.uk, or telephone (+44) 01280 827460

Visit us at Cyto 2015!

The 30th Congress of the International Society for Advancement of Cytometry is taking place this
year at the Scottish Exhibition and Conference Centre in Glasgow on the 27-30th June 2015. The meeting will cover talks from worldwide leaders in the flow cytometry field on the advancement of the science and practice of cytometry.

Come and visit us at booth 420 for a chance to win an iWatch™ and to find out more about our range of flow cytometry products available. If you would like to arrange a meeting with us during the congress, please contact
marketing@caltagmedsystems.co.uk.

For further information about Cyto 2015, please click here.










Friday, 12 June 2015

Renal Cell System


The renal cell system removes waste from the body, regulates electrolyte balance and blood pressure, balances the body’s fluids, and controls red blood cell production. Endothelial cells, epithelial cells, and mesangial cells are vital components required for the normal function of the renal system. Atypical regulation and proliferation of these cells can affect renal function and lead to the development of diseases such as diabetic nephropathy, chronic renal disease, polycystic kidney disease, and renal cancer. View Renal Cell System >>
 Featured Products
Human Renal Mesangial Cells
Product Code: 4200 
- Ideal model for studying mesangial injury and glomerular functions under both physiological and pathophysiological conditions.  
- Each vial contains >5 x 10^5 cells in 1 ml volume. 
- Guaranteed to further expand for 15 population doublings. 
- Recommended medium: Mesangial Cell Medium (Product Code: 4201)
 
Human Renal Glomerular Endothelial Cells
Product Code: 4000
- Isolated from human kidneys.   
- Each vial contains >5 x 10^5 cells in 1 ml volume. 
Guaranteed to expand beyond 15 population doublings. 
Recommended medium: Endothelial Cell Medium (Product Code: 1001)
Human Renal Cortical Epithelial Cells
Product Code: 4110
- Useful in vitro model to study the relationship between REpiC and a variety of renal diseases. 
- Each vial contains >5 x 10^5 cells in 1 ml volume.
 Guaranteed to further expand for 15 population doublings. 
Recommended medium: Epithelial Cell Medium (Product Code: 4101)  
2015 ScienCell Research Laboratories|6076 Corte Del Cedro, Carlsbad, CA 92011|(760) 602-8549
Caltag Medsystems is the UK distributor for ScienCell. For more information please contact:
T: +44(0)1280 827460 | www.caltagmedsystems.co.uk | E: office@caltagmedsystems.co.uk

Monday, 8 June 2015

Chromatin Immunoprecipitation Kits

In the past, chromatin immunoprecipitation assays involved lengthy, labour-intensive procedures, often resulting in a low throughput and less enrichment efficiency. Epigentek’s EpiQuick and ChromaFlash ranges of ChIP kits mark a vast improvement in performance, efficiency, and convenience, providing an invaluable tool for studying in vivo protein-DNA interaction. The most popular Epigentek products are detailed in the table below:




Purpose
Selective immuno-precipitation and enrichment of chromatin fractions from low amount of cells or tissues
Selective immuno-precipitation and enrichment of chromatin fractions from cells
Selective immuno-precipitation and enrichment of chromatin fractions from tissues
Selective immuno-precipitation and enrichment of chromatin fractions from plants
Immuno-precipitate and shear isolated chromatin using magnetic beads in one step
Key Points
Recommended for next-gen sequencing applications
Robust protocol designed specifically for cell input
Robust protocol designed specifically for tissue input
Robust protocol designed specifically for plant tissue input
Ultra-fast protocol designed specifically for isolated chromatin input
Starting Material
Cells or Tissues
Cells
Tissues
Plant Tissues (flowers, leaves, young seedlings)
Isolated Chromatin
Input Range
2,000 to 1,000,000 cells or 0.5 to 50 mg tissue per reaction
1,000,000 to 2,000,000 cells per reaction
10 to 30 mg tissue per reaction
20 to 50 mg tissue per reaction
5 to 10 µg chromatin per reaction
Total Protocol Time
5 hours
5 hours
5 hours
6 hours
1 hour (with EpiSonic) or 4 hours (without EpiSonic)
Relative-Fold Enrichment vs IgG
500+
100 to 200
100 to 200
100 to 200
100 to 200
*All products include negative controls, DNA purification columns, and control primers where required*


ChromaFlash ChIP Kits

Epigentek’s ChromaFlash CHiP kits provide fast, cost efficient immunoprecipitation of specific chromatin fragments. The microplate format allows flexibility, allowing single reactions or high throughput assays, and all products are compatible with various downstream analysis workflows including ChIP-PCR, ChIP-on-chip, and ChIP-seq.

The ChromaFlash High Sensitivity ChIP Kit allows for selective immunoprecipitation and enrichment of chromatin fractions from a low amount of cells or tissues whilst reducing background noise.
Product code: P-2027-24 (24 reactions)| Price: £216.00
Product code: P-2027-48 (48 reactions)| Price: £347.00

The ChromaFlash One-Step Magnetic ChIP Kit is used to immunoprecipitate and shear isolated chromatin using magnetic beads in one step. The ultra-fast protocol is designed specifically for isolated chromatin input.
Product code: P-2026-48 (48 reactions)| Price: £276.00
Product code: P-2026-96 (96 reactions)| Price: £465.00

EpiQuik ChIP Kits

Epigentek’s EpiQuik range provides a rugged procedure for selective immunoprecipitation and enrichment of chromatin fractions from a variety of starting materials. The convenient strip microplate format allows for manual or high throughput assays.

EpiQuik ChIP Kit for chromatin immunoprecipitation from cell input.
Product code: P-2002-1 (24 reactions)| Price: £198.00
Product code: P-2002-2 (48 reactions)| Price: £321.00
Product code: P-2002-3 (96 reactions)| Price: £486.00

EpiQuik Tissue ChIP Kit for chromatin immunoprecipitation from tissue input.
Product code: P-2003-1 (24 reactions)| Price: £198.00
Product code: P-2003-2 (48 reactions)| Price: £321.00
Product code: P-2003-3 (96 reactions)| Price: £486.00

EpiQuik Plant ChIP Kit for chromatin immunoprecipitation from plant tissue input.
Product code: P-2014-24 (24 reactions)| Price: £260.00
Product code: P-2014-48 (48 reactions)| Price: £425.00

Also introducing…

EpiNext ChIP-Seq High-Sensitivity Kit (Illumina)

The EpiNext ChIP-Seq High Sensitivity Kit is designed to streamline the process of ChIP sequencing, combining selective enrichment of a chromatin fraction containing specific DNA sequences and preparation of a ChIP-Seq library for next generation sequencing.
  • Capture of low abundance protein/DNA complexes with minimized non-specific background levels.
  • Construct both non-barcoded (singleplexed) and barcoded (multiplexed) ChIPSeq libraries quickly with reduced bias for use with Illumina sequencing platforms.
Product code: P-2030-12 (12 reactions)| Price: £459
Product code: P-2030-24 (24 reactions)| Price: £858


Caltag Medsystems are the UK distributor for Epigentek. If you would like further information about their ChIP kits or any of our products please contact us at office@caltagmedsystems.co.uk or telephone 01280827460

Evaluation of CD4 T-Cell Functional Responses – Introducing The Act T4 Cell Kit

The Act-T4 Cell™ kit is designed to be used in routine, clinical testing and in research to determine the CD4 T-cell-mediated immune response against pathogens, auto-antigens and allergens. The kit consists of an antibody cocktail which allows the identification and quantification of effector CD4 T-cells after incubation of whole blood with mitogens and/or antigens of interest. The total percentage increase of activated CD4 T-cells is related to the immune response against the stimulus used.

Features of the Act T4 Cell Kit:


- Enables the measurement of non-specific and antigen-specific CD4+ T cells (CD3+, CD4+,       CD134+ and CD25+).
- Allows isolation of activated CD4 T-cells for their application in further studies.
- Provides highly accurate and repeatable results on immunosuppressed and paediatric patients.

Several studies have recently validated the pre-clinical use of the Act-T4 Cell™ kit, to evaluate antigen-specific response against virus (e.g. CMV, EBV, HCV, HIV) or bacteria (e.g. Mycobacterium tuberculosis). Results were consistent with serology testing, cell proliferation studies and other techniques based on IFN-γ production levels, revealing the potential of Act-T4 Cell™ to be an accurate, easy and cost-effective method.



CD25+, CD134+, and CD4+ T Cells in response to mitogens and antigens. a) Dot-plots from a representative healthy adult control showing expression for CD25+, CD134+ and CD4+ T Cells in the control culture (0) compared with staining in response to PHA and SEB. b) Representative dot-plots from healthy adult controls showing expression for CD25+, CD 134+ and CD4+ T Cells in response to MTB, CMV and Candida. The percentage of CD25+, CD134+ and CD4+ T Cells is shown for each dot-plot. The gates for CD25+, CD134+ and CD4+ T Cells were based on comparison of negative control (no antigen) and positive control (PHA).






The Act-T4 Cell™ kit is intended to be used after incubating whole blood during 44-48 hours in a culture medium with the antigens/mitogens of interest. The use of a positive control (mitogens such as phytohaemagluttinin or staphylococcal enteroantigen B) and a negative control (unstimulated whole blood) are highly recommended. The antibody combination proposed enables the selection of activated antigen-specific CD4+ T cells.

The Act-T4 Cell™ kit includes two components with enough volume for 100 tests:
- One vial with the following 4-antibody combination:







- Erythrocyte lyse-non-wash solution (QuicklysisTM)

Product Information:
Product Name: Act-T4 Cell Kit
Product Code: CYT-AT4C
£839 (100 tests)
For further information about the Act-T4 Cell Kit, please click here

If you wish to view the reference values to quantify the increase of activated CD4+ T Cell subsets, please click here to access the paper ‘Establishment of a healthy human range for the whole blood “OX40” assay for the detection of antigen-specific CD4+ T cells by flow cytometry’ – Sadler et al (2014), Cytometry Part B.

Caltag Medsystems are the exclusive UK distributor for Cytognos. If you have any questions about the Act-T4 Cell Kit or any of our products, please do not hesitate to contact us at office@caltagmedsystems.co.uk or telephone (+44) 01280 827460.

Functional Antibodies From Adipogen

Antibodies displaying an agonist or antagonist activity (functional grade antibodies (FuncAbs™)) are powerful tools for mimicking or blocking physiological functions in vitro and in vivo.

The mechanism of action for functional antibodies varies depending on its role:

Blocking: Antibodies can bind directly to ligands, preventing them from activating their cognate receptors or binding directly to their cognate receptors and inhibit receptor activation or function.  A potent blocking antibody is anti-Angiopoietin-2 mAb (rec.), able to inhibit the binding of mouse and human angiopoietin-2 to its receptor Tie-2.

Activation/Induction: Binding to receptors can also induce active signals that alter cellular fates. A potent activating antibody is anti-CD40 (mouse), mAb (FGK45), able to activate B and NK cells in vivo and in vitro.

Depletion: Binding of cell surface receptors can result in depletion of antigen-bearing cells through complement-mediated lysis and opsonisation, as well as Fc receptor for IgG (FcγR)-mediated clearance. A potent neutralizing antibody is anti-Neutrophils (mouse), mAb (blocking) (Nimp-R14), able to deplete 98% neutrophils in vivo (250 μg/mouse).

Functional grade antibodies from AdipoGen Life Sciences are available free of preservatives and tested for low endotoxin content and may be used for activation, neutralizing or blocking studies, both in vitro or in vivo.

We are offering a 10% discount across all functional antibodies from the Adipogen Brochure until the 8th September 2015. Simply quote 'FUNC45' with your order to be eligible for the discount.

Antibodies Available:

Blocking/Neutralising Antibodies: 

anti-APRIL (mouse), mAb (rec.) (blocking) (Apry-1-1)
- Inhibits binding of mouse APRIL to mouse BCMA and TACI

anti-Angiopoietin-2 (human), mAb (rec.) (blocking) (Angy-1-4) 
- Inhibits the binding of human angiopoietin-2 to human Tie-2 (Human IgG2λ)

anti-Angiopoietin-2 mAb (rec.) (blocking) (Angy-2-1)
- Inhibits the binding of mouse/human angiopoietin-2 to human Tie-2 (Mouse IgG2bλ)

anti-IL-33 (mouse), mAb (rec.) (blocking) (Bondy-1-1)
- Inhibits the binding of mouse IL-33 to ST2/IL-1RAcP

anti-BAFF (human), mAb (blocking) (4.62)
- Inhibition/Neutralising of human BAFF binding to Raji cells

anti-TRAIL-R1 (human), mAb (HS101)
- Inhibition/Neutralising (blocks TRAIL-R1 mediated killing if applied in solution)

anti-TRAIL-R2 (human), mAb (HS201)
- Inhibition/Neutralising (blocks TRAIL-R2 mediated killing if applied in solution)

anti-NS5B (HCV), mAb (blocking) (5B-12B7)
- Blocks the RNA-dependent RNA polymerase activity in vitro

anti-BTLA (human), mAb (6F4)
- Inhibits interaction of BTLA to HVEM or UL144

anti-LAG-3, mAb (blocking) (11E3)
- Blocks LAG-3/MHC class II interactions

anti-LAG-3 (human), mAb (blocking) (17B4)
- Blocks LAG-3/MHC class II interactions

anti-VEGF-A (human), mAb (3(6D3))
- Inhibits VEGF-A signaling

Inducing/Activating Antibodies:

anti-LTβR (mouse), mAb (4H8 WH2)
- Agonists inducing BAFF, chemokines and integrins in vitro and in vivo (Rat IgG2a)
- Also suitable for FACS

anti-LTβR (mouse), mAb (3C8)
- Agonists inducing BAFF, chemokines and integrins in vitro and in vivo (Rat IgG1κ)

anti-CD40 (mouse), mAb (FGK45)
- Activates B and NK cells in vivo and in vitro

anti-Fas (human), mAb (APO-1-3)
- Induces apoptosis with or without cross-linking (Protein A), depending on cell type

Depleting Antibodies:

anti-BAFF-R (mouse), mAb (9B9)
- Depletes B cells in vivo

anti-Neutrophils (mouse), mAb (blocking) (Nimp-R14)
- Optimal reagent to deplete neutrophils in vivo (250 μg/mouse)

Conformation-specific Recombinant Antibodies:

anti-Rab1-GTP, mAb (rec.) (ROF7)
- Recognises human, mouse, rat and dog Rab1a-GTP and Rab1b-GTP

anti-Rab6-GTP, mAb (rec.) (AA2)
- Recognises human, mouse and drosophila GTP-bound
Rab6a and Rab6b and mutant Rab6Q72L. Does not detect
Rab6-GDP

anti-Tubulin-GTP, mAb (rec.) (MB11)
- Recognises human, mouse, rat and drosophila tubulin-GTP

Other Recombinant Monoclonal Antibodies:

anti-Giantin, mAb (rec.) (TA10)
- Recognises human and mouse giantin

anti-HMGB1, mAb (rec.) (Giby-1-4)
- Recognises human, mouse and rat HMGB1

anti-IL-1R2 (mouse), mAb (rec.) (Praxy-1-1)
- Recognises mouse IL-1R2

anti-Myosin IIA (non-muscle) (heavy chain), mAb (rec.) (SF9) 
- Recognises human, mouse, rat and drosophila myosin
IIA (heavy chain)

anti-α-Tubulin, mAb (rec.) (F2C)
- Recognises mouse, bovine and human α-tubulin

anti-β-Tubulin, mAb (rec.) (S11B)
- Recognises human, mouse, rat, pig, drosophila and
monkey β-tubulin

anti-IL-33 (mouse), mAb (rec.) (Carly-1-4)
- Recognises mouse IL-33

anti-PEDF (human), mAb (rec.) (Serpy-1-4)
- Recognises human PEDF

anti-EGFP, mAb (rec.) (G3)
- Recognises EGFP, ECFP and EYFP



Post-translational Modification-specific Antibody for
Cancer Research:


anti-Polyglutamylation Modification, mAb (GT335)
- Recognises most forms of polyglutamylated tubulin (α- and β-tubulin), independent of the length of the glutamate side chains. No specificity to particular tubulin isoforms nor to tubulin from particular species are observed. Detects also other (poly)glutamylated proteins. Since no consensus modification site is known for protein (poly)glutamylation, the detection is not sequence-specific. However, an acidic environment of the modification site is required.

Inflammasome Signalling Blocking Antibody:

anti-Asc, pAb (AL177)
- Inhibits interaction between Asc and NLRP3, leading to blockade of caspase-1 processing in vitro.

Caltag Medsystems are the UK distributor for Adipogen Life Sciences. For more information about any of our products, please contact us at office@caltagmedsystems.co.uk or call (+44) 01280 827460.

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