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| 1. |
O'Brien, M.A., Moravec, R.A., and Riss T.L.
(2001)
Poly (ADP-ribose) polymerase cleavage monitored in situ in apoptotic cells.
Biotechniques
30
,
886-891
.
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Notes:
During apoptosis, Poly (ADP-ribose) polymerase (PARP) is cleaved by caspase-3 to generate an 85 kDa fragment (p85). An affinity-purified polyclonal antibody to the p85 fragment of PARP is characterized. In Western blot analysis with Jurkat cells treated with an anti-Fas antibody and with SH-Sy5Y cells treated with staurosporine, the antibody recognizes the 85-kDa (p85) fragment of PARP but not full-length PARP. The antibody was used at a concentration of 0.75 µg/ml for Western blots and at 2.5µg/ml for immunocytochemistry. TUNEL labeling of apoptotic Jurkat cells was performed with the DeadEnd™ Fluorometric TUNEL System. Immunocytochemistry was also performed using the Anti-β III Tubulin mAb. Good experimental details are given.
(0002355) |
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Products: Anti-βIII Tubulin mAb | Anti-PARP p85 Fragment pAb | DeadEnd™ Fluorometric TUNEL System |
| 2. |
Li, X. and Darzynkiewicz, Z.
(2000)
Cleavage of poly(ADP-ribose) polymerase measured in situ in individual cells: Relationship to DNA fragmentation and cell cycle position during apoptosis.
Exp. Cell Res.
255
,
125-132
.
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Notes:
The Anti-PARP p85 Fragment pAb was used in an extensive analysis of PARP cleavage relatively to DNA strand breaks induced in HL-60 cells. Apoptosis mediated by either campothecin or TNFα. PARP cleavage was detected by immunocytochemistry as well as multiparameter flow and laser scanning cytometry. PARP cleavage was readily apparent 30 minutes after TNFα treatment and PARP cleavage was apparent 90 minutes after campothecin treatment. The PARP cleavage was specific to S-phase of the cell cycle in the campothecin treated cells and was not specific to any cell phase during TNFα treatment. DNA strand breaks followed PARP cleavage by about 30 minutes. A strong correlation between DNA strand breaks and PARP cleavage is demonstrated.
(0000801) |
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Products: Anti-PARP p85 Fragment pAb |