DNA Fragmentation Detection Kit (60 slides)
AS20 4459 | The DNA Fragmentation Detection Kit can be used for the identification apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue sections, or in preparations of single cell suspensions fixed on slides.
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Background | This DNA Fragmentation Detection Kit utilizes the fact that Terminal deoxynucleotidyl Transferase (TdT) binds to exposed 3’-OH ends of DNA fragments. During apoptosis, such DNA fragments are generated, which catalyzes the addition of biotin-labeled deoxynucleotides. A streptavidin-horseradish peroxidase (HRP) conjugate is used to detect biotinylated nucleotides, and diaminobenzidine (DAB) reacts with the HRP-labeled sample, generating a
brown, insoluble substrate at the sites where DNA fragmentation has taken place. Counterstaining with Methyl Green allows the morphological evaluation and characterization of normal and apoptotic cells. The DNA Fragmentation Detection Kit can be used for the identification apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue sections, or in preparations of single cell suspensions fixed on slides.
This kit contains the following reagents:
Proteinase K, a pH-stabilized Solution, 100 μl
TdT Equilibration Buffer, 8 ml
TdT Labeling Reaction Mix (mix of labeled and unlabeled nucleotides), 2 x 1.3 ml
TdT Enzyme (Terminal Deoxynucleotidyl Transferase), 70 μl
Stop Buffer, 8 ml
Block Buffer, 24 ml
Streptavidin-HRP Conjugate (25x concentrate), 300 μl
DAB Concentrate, 300 μl
DAB Reaction Buffer, 8 ml
Methyl Green Counterstain, 2 x 3.5 ml
This kit contains reagents for 60 tests.
Storage | Store kit at -20°C. For long-term storage, it is recommended to keep the TdT Enzyme, TdT Labeling Reaction Mix
and Streptavidin-HRP Conjugate (25x concentrate) frozen in aliquots. Use only the amount needed of these
components for each test, while leaving the remaining aliquots at -20°C. For short-term storage (up to 2 weeks),
the kit components can be kept at 4-8°C.
This kit should not be used beyond the expiration date.
Manual in pdf
native papain isolated and purified from Carica papaya
|Purity:||Purified IgG in PBS. Contains 0.08% sodium azide.|
|Reconstitution:||For reconstitution add 0,5 ml of sterile destilled water|
|storage:||Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.|
|Tested applications:||ELISA (ELISA), Immunofluorescence (IF), Immunohistochemistry (IHC), Western blot (WB)|
|recommended dilution:||1 : 1000-1 : 100 000 (ELISA), (IF), (IHC), (WB)|
|Expected | apparent MW:||38,9 kDa|
|Confirmed reactivity:||Carica papaya|
|predicted reactivity:||Carica papaya|
|not reactive in:||No confirmed exceptions from predicted reactivity are currently known|
|additional information:||Antibody is labelled with biotin using N-hydroxysuccinimidobiotin, Antibody potency and purity has been evaluated by immunoelectrophoresis, single radial immunodiffusion (Ouchterlony), ELISA,immunoblotting and enzyme inhibition|
|additional information (application):||Biotin/IgG protein molar ration is approximately 6,2, No foreign proteins are added|
Papain (EC=184.108.40.206) is a cysteine protease which belongs to peptidase C1 family. Can cause an allergic reaction in humans.