Phospho-Bad (S112) and Total Bad ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Bad (S112) and Total Bad in Human in Cell Lysate samples.
Application | Reactivity | Dilution info | Notes |
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Phospho-Bad (S112) and Total Bad ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Bad (S112) and Total Bad in Human in Cell Lysate samples.
Phospho-Bad (S112) and Total Bad ELISA Kit (ab279746) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated Bad protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-Bad and total Bad. An anti-pan Bad antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Bad present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-Bad (S112) antibody is added to detect phosphorylated Bad. In the remaining wells, rabbit anti-pan-Bad antibody is used to detect pan Bad. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Bad (S112) or pan Bad bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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The Bad protein also known as Bcl-2-associated death promoter plays a critical role in apoptosis regulation. It possesses a molecular weight of approximately 23 kDa. Bad is expressed widely in tissues and is a member of the Bcl-2 family of proteins. This family is well-known for its involvement in the regulation of cell death both promoting and inhibiting depending on the protein’s interactions.
Its main role is to promote apoptosis by binding to anti-apoptotic proteins like Bcl-2 and Bcl-XL displacing pro-apoptotic Bax and Bak proteins to trigger cell death. Bad functions as part of the mitochondrial apoptosis pathway and is known to form complexes with phosphorylated proteins. Phosphorylation of Bad by kinases can lead to its sequestration to the cytoplasm reducing apoptosis.
Its action is pivotal in the PI3K/Akt signaling pathway. This pathway involves growth factors and cell survival signals where phosphorylation of Bad inhibits its pro-apoptotic activity. Bad interacts with proteins like Akt and 14-3-3 to modulate cell survival. Disruption in its regulation through these pathways may lead to abnormal cell survival or death.
Bad has implications in cancer and neurodegenerative diseases. Cancer cells often exhibit impaired apoptotic pathways where overexpression of Bad can ameliorate responsiveness to therapy. In neurodegenerative diseases such as Alzheimer's dysregulation of its function can contribute to cell death. The protein’s interaction with Bcl-2 and Bcl-XL is significant in these contexts influencing disease propagation and severity.
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Positive Control.
HeLa cells were treated with Calyculin A.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
HeLa cells were untreated or treated with Calyculin A.
HeLa cells were untreated or treated with Calyculin A.
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