Thursday, 19 February 2015

Leukapheresis Packs: Mononuclear Cells and Platelet Rich Plasma

I want to make you aware that AllCells can provide Leukapheresis Packs (LeukoPaks).

Leukapheresis Packs available from AllCells:
LeukoPaks: Mononuclear Cells and Plasma
Mobilised LeukoPaks: Ideal for large scale CD34+ Isolations
T Cell Isolation Options 

LeukoPaks: Mononuclear Cells and Plasma

Normal Healthy Leukapheresis Packs (LeukoPaks) are available in different sizes and with flexible collection options. In all LeukoPaks, the donors total blood volume is circulated through Cobe Spectra machines.

Full LeukoPak contains 150ml of mononuclear cells and 150ml of autologous platelet rich plasma. Half and Quarter LeukoPaks which contain between 50-75ml of mononuclear cells plus 50-75ml of autologous platelet rich plasma are also available. 

Full Pack:  Average Cell Number: 1.5 x 1010  Average Range: 1 to 3.5 x 1010  
Product Code: PB007

Half Pack:  Average Cell Number: 0.75 x 1010  Average Range: 0.6 to 1 x 1010  
Product Code: PB007-1

Quarter Pack:  AverageCell Number: 0.37 x 1010  Average Range: 0.1 to 0.71010  
Product Code: PB007-2

If you require Leukapheresis products from the same donor over a short period of time, AllCells can arrange for a Dual Half Pak collection.  This allows them to collect a Half Pack from the donor and then reserve the donor for a second Half Pack collection 6-12 weeks later.  Once a Full Leuko Pak volume has been collected, the donor must wait 6 months to undergo Leukapheresis again. 

Mobilised LeukoPaks: Ideal for large scale CD34+ Isolations

For customers looking to perform large scale CD34+ cell isolations, AllCells recommend Mobilised LeukoPaks.  Mobilisation involves a series of injections of G-CSF or one injection of Mozobil to the donor, this allows the donors to secrete the rich lymphocyte cell populations found in the bone marrow into the blood stream. A Leukapheresis collection is then performed.  Donors can perform a maximum of two mobilisations in their lifetime, with a one year period in between.  Only full packs are available for mobilised LeukoPaks.

AllCells have 4 mobilisation regiment options that our customers can choose from:

Product Code
Regiment
Quantity
Mozobil @20 mg/ml, 1.2ml single dose – Regimen A, apheresis 9-11 hours post injection  
1 Full pack
G-CSF @7.5-9.6 ug/kg/day x 3 days – Regimen A, apheresis on day 4           
1 Full pack
G-CSF @5 ug/kg/day x 5 days – Regimen E, apheresis on day 5                              
1 Full pack
G-CSF @6.9-9 ug/kg/day – Regimen C, apheresis on days 5 and 6
2 Full packs

Please note that each donor’s reaction to mobilisation is different, resulting in mobilised LeukoPaks with very high total cellularity count and some with lower total cellularity count per pack.  For best CD34+ isolation results, AllCells recommend mPB025-Reg.E.

T Cell Activation

For those customers looking at T-cell activation, AllCells recommend obtaining Whole Blood from a few donors, isolating the T-cells (which they can do for you) and testing for activation. AllCells will reserve the donors until your initial testing is complete. Let us know which donor worked best for your research and the donor will be recalled for a Full LeukoPak collection. 

Shipment of LeukoPaks

AllCells ship their fresh Leuko Paks on cold gel packs to customers in the UK and Ireland.  This slows the metabolism of the cells down for the overseas journey.

Please do not hesitate to contact Caltag Medsystems, the exclusive distributor of AllCells products in the UK, for further information. Email office@caltagmedsystems.co.uk, call +44 (0)1280 827460 or visit our website www.caltagmedsystems.co.uk.

Wednesday, 18 February 2015

Reporter Gene Stabilised mRNA

Reporter Gene mRNAs

Reporter genes are commonly used in cell biology research. Reporter gene mRNA can be used as controls to study transfection and expression in mammalian cells using a variety of assays. The capped (Cap 0) and polyadenylated mRNAs is optimised for mammalian systems and modified to reduce immune stimulation. They mimic fully processed mature mRNA.

 mRNA transfection provides several advantages :

             -No need for nuclear uptake - protein expression occurs directly in cytoplasm
     -Faster protein expression than DNA transfection
     -No genomic integration
     -Protein expression directly related to mRNA quantity
     - Perfect for transfecting slow or non-dividing cells
     - Protein expression in a total promoter-independent manner
     -Transient transfection: mRNA based expression of proteins is sustained for a limited time

GFP mRNA has been designed to express high levels of Green Fluorescent Protein. It is a commonly used direct detection reporter in mammalian cell culture, yielding bright green fluorescence with an emission peak at 509 nm. 













LUC mRNA
has been designed to express high levels of FireFly Luciferase. It is commonly used in mammalian cell culture to measure both gene expression and cell viability. It emits bioluminescence in the presence of the substrate, luciferin.

mCherry mRNA encodes the fluorescent protein, mCherry, which is derived from DsRed, a protein found in Discosoma sp. mCherry is a monomeric fluorophore with a peak absorption at 587 nm and emission at 610 nm. It is photostable and resistant to photobleaching. 

β-galactosidase (β-gal) mRNA, codes for a protein product of the bacterial LacZ gene. β-gal catalyses the conversion of β-galactosides into monosaccharides. It is a common marker gene used to assess transfection efficiency.

We are offering a 10% discount on the following products in the table below. Hurry though, as this offer ends on the 27th February 2015.

Product Code
Description
Size
mRNA GFP
20 µg
mRNA GFP
100µg
mRNA Luciferase
20µg
mRNA Luciferase
100µg
mCherry mRNA
20µg
mCherry mRNA
100µg
Beta Gal mRNA
20µg
Beta Gal mRNA
100µg



Caltag Medsystems are the UK distributor for OZ Biosciences. For further information about their products, please visit our website, www.caltagmedsystems.co.uk, call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

Tuesday, 17 February 2015

Genome Editing Stabilised mRNA

Genome Editing mRNAs

Plasmids and viral vectors are traditionally used in CRISPR genome editing to express the required proteins. However, there is a risk in using DNA instead of mRNA: Double stranded DNA breaks catalyse insertion of DNA at the cut site. At some substantial frequency, the protein expression vectors can integrate which can lead to continuous expression of the nuclease or a previously silent sequence.

Now mRNA is becoming a powerful tool in genome editing. With no risk of insertional mutagenesis, mRNA can be used to transiently express the required proteins. Wild-type Cas9 mRNA is stabilised non-immunogenic messenger RNA (mRNA) that has been designed to induce a high level of expression of wild-type Cas9 protein. This creates a double stranded break at a target site delineated by RNA guide sequences.

This mRNA is produced by in vitro transcription, stabilised at the 5’ end by modified nucleotides capping and contain a poly(A) tail at the 3’ end. Cas9 mRNA has been optimised for improved stability, improved performance, and to lower the cellular innate immunity response.

mRNA transfection provides several advantages:

-No need for nuclear uptake - protein expression occurs directly in cytoplasm
-Faster protein expression than DNA transfection
-No genomic integration
-Perfect for transfecting slow or non-dividing cells
-Protein expression in a total promoter-independent manner
-Transient transfection: mRNA based expression of proteins is sustained for a limited time.

We are offering a 10% discount on the following products in the table below. Hurry though, as this offer ends on the 27th February 2015.

Product Code
Description
Size
Cas9 mRNA
20µg
Cas9 mRNA
100µg
CRE mRNA
20µg
CRE mRNA
100µg


Caltag Medsystems are the UK distributor for OZ Bioscience. For further information about their products, please visit our website, www.caltagmedsystems.co.uk, call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

Vaccine Stabilised mRNA

Vaccine stabilised mRNA offers several advantages over traditional DNA and viral-based approaches:

-Express the desired antigen and act as an adjuvant.
-Prevent risk of genomic integration and no inactivated viruses or pathogens are needed.
-No need for nuclear uptake - protein expression occurs directly in cytoplasm
-Perfect for transfecting slow or non-dividing cells, such as Antigen Presenting cells (DCs)
-Faster protein expression than DNA transfection
-Protein expression in a total promoter-independent manner
-Transient transfection: mRNA based expression of proteins is sustained for a limited time.

We offer modified OVA mRNA and unmodified OVA mRNA:

-Exogenous unmodified mRNAs activate the innate immune system and production of cytokines in order to influence an induced immune response, but is rapidly degraded.
-Modified mRNAs reduce this effect but last longer in the organism due to their improved stability and additional and protections.

The capped (Cap 0) and polyadenylated mRNA is optimised for mammalian systems with modified nucleotides to reduce immune stimulation.

We are offering a 10% discount on the following products in the table below - hurry though as this offer ends on the 27th February 2015.


Product Information:

Product Code
Description
Size
Modified OVA mRNA
20µg
Modified OVA mRNA
100µg
Unmodified OVA mRNA
20µg
Unmodified OVA mRNA
100µg
Beta Gal mRNA
20µg
Beta Gal mRNA
100µg



Caltag Medsystems are the UK distributor for OZ Bioscience. For further information about their products, please visit our website, www.caltagmedsystems.co.uk, call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

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