Phospho-MCL-1 (Ser159), PE, clone: RBCERNR, eBioscience™

Description

Mcl-1 (Myeloid cell leukemia-1) is a member of the bcl-2 family. The carboxy terminal of Mcl-1 and bcl-2 share significant sequence homology. Expression of Mcl-1 is increased upon exposure of ML-1 cells to various types of DNA damaging agents (e.g. ionizing radiation, ultraviolet radiation, and alkylating drugs) along with increases in GADD45 and Bax and a decrease in bcl-2. Enhanced expression of Mcl-1, prominently associated with mitochondria, complements the continued expression of bcl-2 in ML-1 cells undergoing differentiation. Like bcl-2, Mcl-1 has the capacity to promote cell viability under conditions that otherwise cause apoptosis.

Description: This RBCERNR monoclonal antibody recognizes human and mouse myeloid cell leukemia sequence 1 (Mcl-1) when phosphorylated on serine 159 (S159). Mcl-1 is an anti-apoptotic protein that is a member of the Bcl-2 family of proteins important for regulation of cell survival/apoptosis. Mcl-1 is primarily localized to the outer membrane of mitochondria where it prevents cytochrome c release via dimerization with other Bcl-2 family members such as Bim. PI3K activation of AKT results in the phosphorylation of GSK3 beta at serine 9 (S9) resulting in destabilization and degradation of GSK3 beta. Loss of GSK3 beta prevents phosphorylation of Mcl-1 on S159 and its subsequent ubiquitnation and degradation. Mice conditionally lacking Mcl-1 in lymphocytes showed that Mcl-1 is essential during early lymphoid development and for the maintenance of mature lymphocytes.Applications Reported: This RBCERNR antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This RBCERNR antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.125 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.Staining Protocol: Protocol A and Protocol C are recommended for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to Clone Performance Following Fixation/Permeabilization located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: Staining Intracellular Antigens for Flow Cytometry Protocol located in the Best Protocols Section under the Resources tab online.Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2μm post-manufacturing filtered.