Phospho-Rb (T826) and Total Rb ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Rb (T826) and Total Rb in Human in Cell Lysate samples.
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Phospho-Rb (T826) and Total Rb ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-Rb (T826) and Total Rb in Human in Cell Lysate samples.
Phospho-Rb (T826) and Total Rb ELISA Kit (ab279906) 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 Rb 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-Rb and total Rb. An anti-pan Rb antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Rb present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho Rb (T826) antibody is added to detect phosphorylated Rb. In the remaining wells, rabbit anti-pan-Rb antibody is used to detect pan Rb. 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 Rb (T826) or pan Rb 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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Retinoblastoma protein (Rb) also known as pRb is an important regulatory protein with a molecular weight of approximately 110 kDa. It is mainly expressed in the nucleus of cells. Rb functions as a tumor suppressor by controlling the cell cycle progression from G1 to S phase. Rb becomes active when it is dephosphorylated allowing it to bind and inhibit E2F transcription factors consequently preventing the transcription of genes essential for S phase entry.
Retinoblastoma protein influences cellular proliferation differentiation and apoptosis. It acts within a larger protein complex modulating various cellular responses. When functional Rb halts uncontrolled cell division important for maintaining normal tissue homeostasis. In its phosphorylated state known as phospho-Rb or phospho-Rb E182 it loses its regulatory capabilities which can lead to unrestrained cell cycle progression.
Several involve the retinoblastoma protein. One key pathway is the p53 pathway which Rb interacts with to influence cellular outcomes like cell-cycle arrest and apoptosis. Rb cooperates with proteins like p21 to implement these processes. Additionally it is involved in the cyclin D-dependent kinase 4 (CDK4) pathway which modulates its phosphorylation state influencing binding interactions and cell cycle control.
Dysregulation of retinoblastoma protein is commonly associated with cancer particularly retinoblastoma and breast cancer. In retinoblastoma mutations in the Rb gene directly lead to uncontrolled cell proliferation due to the absence of functional Rb protein. Additionally Rb's connection to the E2F family of transcription factors can become disrupted contributing to oncogenesis in other cancers.
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Positive Control.
HT29 cells were treated with Thymidine and Nocodazole.
Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.
HT29 cells treated with/without Thymidine-Nocodazole Block. HT29 cells were treated with Thymidine-Nocodazole Block
HT29 cells treated with/without Thymidine-Nocodazole Block. HT29 cells were treated with Thymidine-Nocodazole Block
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