ELISPOT and FLUOROSPOT kits

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1 ELISPOT and FLUOROSPOT kits Interleukins Interferons Granzymes and perforins TNF superfamily ligands and receptors Apoptosis markers And many more...

2 ELISPOT and FLUOROSPOT: a cell-based assay to assess immune response The ELISPOT assay is a highly sensitive technique facilitating the detection of individual cells secreting a particular target protein, in response to various drugs, stimuli or inhibitors. ELISPOT is often used to investigate the Th1/Th2 response, vaccine development, viral infection, infectious diseases, autoimmune diseases and transplantation, among others. FLUOROSPOT offers the same application, but with the added flexibility of fluorescence detection. A dual detection assay allows the observation of the relationship between the expression of two proteins. ELISPOT kit components Capture antibody or pre-coated 96 well plates (varies by kit format*) Biotinylated detection antibody Streptavidin-Alkaline Phosphatase conjugate Ready-to-use substrate buffer Bovine Serum Albumin Dry skimmed milk Dual ELISPOT kit components 96 PVDF-bottomed-well plates* Capture antibody for protein 1 Capture antibody for protein 2 Biotinylated detection antibody for protein 1 FITC conjugated detection antibody for protein 2 Anti-FITC antibody HRP conjugate Streptavidin-Alkaline Phosphatase conjugate Bovine Serum Albumin Dry skimmed milk Ready-to-use AEC substrate buffer (red spots) Ready-to-use BCIP/NBT substrate buffer (blue spots) Dual FLUOROSPOT kit components 96 PVDF-bottomed-well plates* Capture antibody for protein 1 Capture antibody for protein 2 Biotinylated detection antibody for protein 1 FITC conjugated detection antibody for protein 2 Anti-FITC antibody green fluorescence conjugate Streptavidin-Phycoerythrin conjugate Bovine Serum Albumin Dry skimmed milk *Some kit formats require the purchase of 96 PVDF-bottomed-well plates separately.

3 Dual ELISPOT Streptavidin- Alkaline Phosphatase mab detection-biotin BCIP/ NBT AP AEC HRP pab anti-fitc - Horse Radish Peroxydase mab detection-fitc Cytokine 1 Cytokine 2 mab capture 1 mab capture 2 Double secreting cells will be observed with purple spots. Dual FLUOROSPOT UV Fluo Fluo AF pab anti-fitc Alexa Fluor Streptavidin-PE mab detection-biotin mab detection-fitc Cytokine 1 Cytokine 2 mab capture 1 mab capture 2 Cells are incubated in 96 well plates on a membrane previously coated with primary antibodies (mab capture 1 and 2). Proteins secreted by these cells bind to the closest primary antibody (proteins 1 and 2). Application of the detection antibodies and subsequent development will result in a colored (ELISPOT) or fluorescent (FLUOROSPOT) spot at the site of the secreting cell, the Spot Forming Cell (SFC; where one spot = one cell).

4 ELISPOT kits from Abcam Target* Reagents for Plates Species Catalog# 96-well plates included reactivity Fas Ligand 5 3 Human ab46605 Fas Ligand 5 Human ab46577 Granzyme B 5 3 Human ab46617 Granzyme B 5 Human ab46581 IL1 Beta 5 3 Human ab46608 IL1 Beta 5 Human ab46578 IL2 10 Mouse ab46625 IL Mouse ab46626 IL2 5 3 Human ab46554 IL2 5 Human ab46546 IL2 5 3 Mouse ab46624 IL2 5 Mouse ab46623 IL4 5 3 Human ab46557 IL4 5 Human ab46547 IL5 5 3 Human ab46611 IL5 5 Human ab46579 IL6 5 3 Human ab46560 IL6 5 Human ab46548 IL Human ab46563 IL10 5 Human ab46549 IL12 p Human ab46574 IL12 p70 5 Human ab46552 IL Human ab46614 IL13 5 Human ab46580 Interferon Gamma 10 3 Mouse ab46589 Interferon Gamma 10 Mouse ab46588 Interferon Gamma 10 Rat ab46592 Interferon Gamma 10 3 Rat ab46593 Interferon Gamma 5 Rat ab46590 Interferon Gamma 5 3 Human ab46569 Interferon Gamma 5 3 Mouse ab46587 Interferon Gamma 5 Mouse ab46586 Interferon Gamma 5 3 Rat ab46591 Perforin 5 Human ab46582 Perforin 5 3 Human ab46620 TNF Alpha 10 3 Rat ab46597 TNF Alpha 10 Rat ab46596 TNF Alpha 5 3 Human ab46566 TNF Alpha 5 Human ab46550 TNF Alpha 5 3 Rat ab46595 TNF Alpha 5 Rat ab46594

5 Dual ELISPOT kits from Abcam Target 1* Target 2 Reagents for Plates Species Catalog# 96-well plates included reactivity Granzyme B Interferon Gamma 5 Human ab48463 Granzyme B Interferon Gamma 5 3 Human ab48464 Granzyme B Interferon Gamma 10 Human ab48465 Granzyme B Interferon Gamma 10 3 Human ab48466 Granzyme B Interferon Gamma 15 Human ab48763 Granzyme B Interferon Gamma 20 Human ab48764 IL2 Interferon Gamma 5 Human ab48447 IL2 Interferon Gamma 5 3 Human ab48448 IL2 Interferon Gamma 10 Human ab48449 IL2 Interferon Gamma 10 3 Human ab48450 IL2 Interferon Gamma 15 Human ab48753 IL2 Interferon Gamma 20 Human ab48754 IL2 IL10 5 Human ab48455 IL2 IL Human ab48456 IL2 IL10 10 Human ab48457 IL2 IL Human ab48458 IL2 IL10 15 Human ab48759 IL2 IL10 20 Human ab48760 IL4 Interferon Gamma 5 Human ab46648 IL4 Interferon Gamma 5 3 Human ab46649 IL4 Interferon Gamma 10 Human ab46650 IL4 Interferon Gamma 10 3 Human ab46651 IL4 Interferon Gamma 15 Human ab48726 IL4 Interferon Gamma 20 Human ab48735 IL4 IL10 5 Human ab46664 IL4 IL Human ab48426 IL4 IL10 10 Human ab48428 IL4 IL Human ab48429 IL4 IL10 15 Human ab48748 IL4 IL10 20 Human ab48749 IL5 Interferon Gamma 5 Human ab46655 IL5 Interferon Gamma 5 3 Human ab46658 IL5 Interferon Gamma 10 Human ab46659 IL5 Interferon Gamma 10 3 Human ab46660 IL5 Interferon Gamma 15 Human ab48743 IL5 Interferon Gamma 20 Human ab48744 IL10 Interferon Gamma 5 Human ab46629 IL10 Interferon Gamma 5 3 Human ab46642 IL10 Interferon Gamma 10 Human ab46643 IL10 Interferon Gamma 10 3 Human ab46644 IL10 Interferon Gamma 15 Human ab48721 IL10 Interferon Gamma 20 Human ab48722 *Many more targets and kit sizes are available at Copyright 2011 Abcam, All Rights Reserved. The Abcam logo is a registered trademark. All information / detail is correct at time of going to print.

6 Dual FLUOROSPOT kits from Abcam Target 1* Target 2 Reagents for Plates Species Catalog# 96-well plates included reactivity Granzyme B Interferon Gamma 5 Human ab48467 Granzyme B Interferon Gamma 5 3 Human ab48468 Granzyme B Interferon Gamma 10 Human ab48469 Granzyme B Interferon Gamma 15 Human ab48765 Granzyme B Interferon Gamma 20 Human ab48766 IL2 Interferon Gamma 5 Human ab48451 IL2 Interferon Gamma 5 3 Human ab48452 IL2 Interferon Gamma 10 Human ab48453 IL2 Interferon Gamma 10 3 Human ab48454 IL2 Interferon Gamma 15 Human ab48756 IL2 Interferon Gamma 20 Human ab48758 IL2 IL10 5 Human ab48459 IL2 IL Human ab48460 IL2 IL10 10 Human ab48461 IL2 IL Human ab48462 IL2 IL10 15 Human ab48761 IL2 IL10 20 Human ab48762 IL4 Interferon Gamma 5 Human ab46640 IL4 Interferon Gamma 5 3 Human ab46645 IL4 Interferon Gamma 10 Human ab46646 IL4 Interferon Gamma 10 3 Human ab46647 IL4 Interferon Gamma 15 3 Human ab48723 IL4 Interferon Gamma 20 Human ab48724 IL4 IL10 5 Human ab46656 IL4 IL Human ab46661 IL4 IL10 10 Human ab46662 IL4 IL Human ab46663 IL4 IL10 15 Human ab48745 IL4 IL10 20 Human ab48746 IL5 Interferon Gamma 5 Human ab48430 IL5 Interferon Gamma 5 3 Human ab48431 IL5 Interferon Gamma 10 Human ab48432 IL5 Interferon Gamma 10 3 Human ab48433 IL5 Interferon Gamma 15 Human ab48751 IL5 Interferon Gamma 20 Human ab48752 IL10 Interferon Gamma 5 3 Human ab46652 IL10 Interferon Gamma 10 Human ab46653 IL10 Interferon Gamma 10 3 Human ab46654 IL10 Interferon Gamma 15 Human ab48741 IL10 Interferon Gamma 20 Human ab48742 *Many more targets and kit sizes are available at 152_11_CT

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