Mouse Cell Death ELISA Kits
Mouse Bcl-2 (B-Cell Leukemia/Lymphoma 2) ELISA Kit (MOES00758)
- SKU:
- MOES00758
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P10417
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- Bcl2, PPP1R50
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Death
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse Bcl-2 in samples. No significant cross-reactivity or interference between Mouse Bcl-2 and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse Bcl-2. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse Bcl-2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse Bcl-2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse Bcl-2. The concentration of Mouse Bcl-2 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | Bcl-2: a antiapoptotic member of the Bcl-2 family. Regulates cell death by controlling the mitochondrial membrane permeability. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). Phosphorylation by JNKs may increase its antiapoptotic functions. |
| UniProt Protein Details: | Protein type:Oncoprotein; Autophagy; Apoptosis; Membrane protein, integral Cellular Component: cytoplasm; cytosol; endoplasmic reticulum; endoplasmic reticulum membrane; integral to membrane; intracellular; membrane; mitochondrial membrane; mitochondrial outer membrane; mitochondrion; myelin sheath; nuclear membrane; nucleus; perinuclear region of cytoplasm; pore complex Molecular Function:BH domain binding; BH3 domain binding; channel activity; channel inhibitor activity; identical protein binding; protease binding; protein binding; protein heterodimerization activity; protein homodimerization activity; protein phosphatase 2A binding; protein phosphatase binding; transcription factor binding; ubiquitin protein ligase binding Biological Process: actin filament organization; apoptosis; apoptotic mitochondrial changes; axon regeneration; axonogenesis; B cell differentiation; B cell homeostasis; B cell lineage commitment; B cell proliferation; B cell receptor signaling pathway; behavioral fear response; CD8-positive, alpha-beta T cell lineage commitment; cell aging; cell growth; cell morphogenesis; cell proliferation; cell-cell adhesion; cellular calcium ion homeostasis; cellular response to glucose starvation; cochlear nucleus development; defense response to virus; developmental growth; digestive tract morphogenesis; DNA damage response, signal transduction resulting in induction of apoptosis; ear development; endoplasmic reticulum calcium ion homeostasis; focal adhesion formation; gland morphogenesis; glomerulus development; growth; hair follicle morphogenesis; hemopoiesis; homeostasis of number of cells within a tissue; immune system development; induction of apoptosis by oxidative stress; induction of apoptosis via death domain receptors; kidney development; leukocyte homeostasis; lymphocyte homeostasis; lymphoid progenitor cell differentiation; male gonad development; melanin metabolic process; melanocyte differentiation; mesenchymal cell development; metanephros development; negative regulation of apoptosis; negative regulation of autophagy; negative regulation of cell growth; negative regulation of cell migration; negative regulation of cell proliferation; negative regulation of cellular pH reduction; negative regulation of mitotic cell cycle; negative regulation of myeloid cell apoptosis; negative regulation of neuron apoptosis; negative regulation of ossification; negative regulation of osteoblast proliferation; negative regulation of retinal cell programmed cell death; neuron apoptosis; oocyte development; organ growth; organ morphogenesis; ossification; ovarian follicle development; peptidyl-serine phosphorylation; peptidyl-threonine phosphorylation; pigment granule organization and biogenesis; pigmentation; pigmentation during development; positive regulation of B cell proliferation; positive regulation of catalytic activity; positive regulation of cell growth; positive regulation of cell proliferation; positive regulation of melanocyte differentiation; positive regulation of multicellular organism growth; positive regulation of neuron maturation; positive regulation of peptidyl-serine phosphorylation; positive regulation of pigmentation; positive regulation of skeletal muscle fiber development; positive regulation of smooth muscle cell migration; post-embryonic development; protein amino acid dephosphorylation; protein polyubiquitination; regulation of apoptosis; regulation of autophagy; regulation of calcium ion transport; regulation of catalytic activity; regulation of cell cycle; regulation of cell-matrix adhesion; regulation of gene expression; regulation of mitochondrial membrane permeability; regulation of mitochondrial membrane potential; regulation of nitrogen utilization; regulation of pigmentation during development; regulation of programmed cell death; regulation of protein heterodimerization activity; regulation of protein homodimerization activity; regulation of protein localization; regulation of protein stability; regulation of transmembrane transporter activity; regulation of viral genome replication; release of cytochrome c from mitochondria; renal system process; response to acid; response to cytokine stimulus; response to DNA damage stimulus; response to drug; response to gamma radiation; response to glucocorticoid stimulus; response to hydrogen peroxide; response to iron ion; response to nicotine; response to oxidative stress; response to steroid hormone stimulus; response to toxin; response to UV-B; spleen development; T cell differentiation; T cell differentiation in the thymus; T cell homeostasis; T cell lineage commitment; thymus development; transmembrane transport; ureteric bud branching; ureteric bud development |
| NCBI Summary: | This gene encodes a member of the B cell lymphoma 2 protein family. Members of this family regulate cell death in multiple cell types and can have either proapoptotic or antiapoptotic activities. The protein encoded by this gene inhibits mitochondrial-mediated apoptosis. This protein is an integral outer mitochondrial membrane protein that functions as part of signaling pathway that controls mitochondrial permeability in response to apoptotic stimuli. This protein may also play a role in neuron cell survival and autophagy. Abnormal expression and chromosomal translocations of this gene are associated with cancer progression in numerous tissues. Alternate splicing results in multiple transcript variants. [provided by RefSeq, Sep 2015] |
| UniProt Code: | P10417 |
| NCBI GenInfo Identifier: | 408360316 |
| NCBI Gene ID: | 12043 |
| NCBI Accession: | P10417. 3 |
| UniProt Secondary Accession: | P10417,P10418, Q4VBF6, |
| UniProt Related Accession: | P10417 |
| Molecular Weight: | 22,281 Da |
| NCBI Full Name: | Apoptosis regulator Bcl-2 |
| NCBI Synonym Full Names: | B cell leukemia/lymphoma 2 |
| NCBI Official Symbol: | Bcl2 |
| NCBI Official Synonym Symbols: | Bcl-2; AW986256; C430015F12Rik; D630044D05Rik; D830018M01Rik |
| NCBI Protein Information: | apoptosis regulator Bcl-2 |
| UniProt Protein Name: | Apoptosis regulator Bcl-2 |
| Protein Family: | Bcl2-associated agonist of cell death |
| UniProt Gene Name: | Bcl2 |
| UniProt Entry Name: | BCL2_MOUSE |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | O.D | Average | Corrected |
| 1000 | 2.362 2.398 | 2.38 | 2.296 |
| 500 | 1.708 1.712 | 1.71 | 1.626 |
| 250 | 1.001 0.987 | 0.994 | 0.91 |
| 125 | 0.516 0.52 | 0.518 | 0.434 |
| 62.5 | 0.305 0.295 | 0.3 | 0.216 |
| 31.25 | 0.193 0.181 | 0.187 | 0.103 |
| 15.63 | 0.131 0.143 | 0.137 | 0.053 |
| 0 | 0.077 0.091 | 0.084 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse Bcl-2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse Bcl-2 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 54.80 | 140.60 | 367.20 | 50.30 | 129.40 | 348.40 |
| Standard deviation | 3.20 | 6.50 | 18.00 | 3.10 | 7.40 | 18.50 |
| C V (%) | 5.84 | 4.62 | 4.90 | 6.16 | 5.72 | 5.31 |
Recovery
The recovery of Mouse Bcl-2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 85-99 | 91 |
| EDTA plasma (n=5) | 95-108 | 101 |
| Cell culture media (n=5) | 88-101 | 94 |
Linearity
Samples were spiked with high concentrations of Mouse Bcl-2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 92-107 | 85-97 | 97-111 |
| Average (%) | 98 | 92 | 105 | |
| 1:4 | Range (%) | 102-119 | 81-95 | 98-112 |
| Average (%) | 108 | 88 | 105 | |
| 1:8 | Range (%) | 96-109 | 86-96 | 97-112 |
| Average (%) | 102 | 91 | 104 | |
| 1:16 | Range (%) | 93-107 | 86-97 | 94-108 |
| Average (%) | 101 | 91 | 99 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
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