Apoptosis PCR Bax/Bcl-2 Multiplex Primer Sets 

Bcl-2 is a member of a large gene family encoding proteins that can either inhibit (e.g. Bcl-2, Bcl-x_L) or promote (e.g. Bax, Bcl-x_S, Bak) apoptosis. Bcl-2 inhibits apoptosis by preventing the release of apoptosis inducing factor (AIF) and cytochrome c from mitochondria. If the Bcl-2 level is higher than the Bax level, apoptosis will be prohibited. The ratio of Bcl-2 to Bax in the cell can determine whether or not the cell initiates apoptosis or survives. Some cancer cells such as melanomas overexpress Bcl-2 preventing apoptosis and allowing malignant growth to continue.

• Simultaneous RT-PCR amplification and detection of Bax and Bcl-2 mRNA levels.
• Includes primers for the GAPDH gene as an internal control for comparison of Bax and Bcl-2 mRNA levels to allow for semiquantitative evaluation of Bax and Bcl-2.
• Functions with Taq DNA polymerase, AccuTaq LA DNA polymerase, REDTaq DNA polymerase, and equivalent DNA polymerases.
• Primer solutions contain both sense and anti-sense primers of the specific gene for convenience.
• The primers work with cDNA reverse transcribed from RNA (mRNA and total RNA) extracted from cells and tissues using various extraction methods.
• Works in one step RT-PCR procedures.
• Genomic DNA contamination in the RNA will not be amplified.

The kit can be used for studying the progress of apoptosis as well as the evaluation of potential apoptosis inducers. It contains primer sets for the detection of Bax and Bcl reverse transcribed from mRNA from rat, mouse, and human tissues or cell lines (reverse transcriptase and thermostable DNA polymerase not included). A primer set for a "housekeeping" gene, GAPDH, as an internal control is included in the kit. These primer sets are optimized for the simultaneous amplification, detection and comparison of Bax and Bcl-2 mRNA levels. The size of the amplified products are 487bp for Bax, 127bp for Bcl-2 and 349bp for GAPDH.

sufficient for 50 50-μl PCR reactions

suitable for mRNA or total RNA extracted from cells and tissues

wet ice

−20°C

Korsmeyer, S.J., Bcl-2 gene family and the regulation of programmed cell death. Cancer Res. 59, 1693s-1700s, (1999) abstract

Oltvai, Z.N., et al., Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74, 609-619, (1993) abstract

Ogawa, N., et al., Lymphocyte apoptosis and apoptosis-associated gene expression in Sjogren's syndrome. Arthritis Rheum. 39, 1875-1885, (1996) abstract

RTN10, GenElute^™ Mammalian Total RNA Miniprep Kit

RTN70, GenElute^™ Mammalian Total RNA Miniprep Kit

D1806, Taq DNA Polymerase from Thermus aquaticus

RTN350, GenElute^™ Mammalian Total RNA Miniprep Kit

STR1, Enhanced Avian First Strand Synthesis Kit

D4545, Taq DNA Polymerase from Thermus aquaticus

T9424, TRI Reagent^®

D7295, Deoxynucleotide Mix, 10 mM

D4309, REDTaq^® DNA Polymerase

MRN70, GenElute^™ mRNA Miniprep Kit

MRN10, GenElute^™ mRNA Miniprep Kit

Bax PCR primer set B8304

Bcl-2 PCR primer set B9179

Control cDNA

GAPDH PCR primer set