Human Total Bcl-2 DuoSet IC ELISA Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4 hours 40 minutes (after plate preparation) | Sample Type & Volume Required Per Well | Cell lysates (100 μL) | Sensitivity | 0.051 ng/mL | Assay Range | 62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva) | Specificity | to measure total Bcl-2 in cell lysates | Cross-reactivity | < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested | Interference | No significant interference observed with available related molecules. |
Product Summary This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total Bcl-2 in cell lysates. An immobilized capture antibody specific for Bcl-2 binds both phosphorylated and unphosphorylated Bcl-2. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: Bcl-2Bcl-2 is a member of a family of proteins that regulates outer mitochondrial membrane permeability. Bcl-2 is an anti-apoptotic member that prevents release of cytochrome c from the mitochondria intermembrane space into the cytosol. Bcl-2 is present on the outer mitochondrial membrane and is also found on other membranes in some cell types. Natural Bcl-2 contains a carboxyl-terminal mitochondria targeting sequence. Long Name: | B Cell Lymphoma/Leukemia 2 | Entrez Gene IDs: | 596 (Human); 12043 (Mouse); 24224 (Rat) | Alternate Names: | apoptosis regulator Bcl-2; B-cell CLL/lymphoma 2; Bcl2; Bcl-2 |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
