Human Bcl-2 ELISA Kit (ab272102)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 38.08 pg/ml
- Range: 195.31 pg/ml - 12500 pg/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Bcl-2 ELISA Kit
See all Bcl-2 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Extract 5 6.3% Inter-assay Sample n Mean SD CV% Extract 3 6.2% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
38.08 pg/ml -
Range
195.31 pg/ml - 12500 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture extracts 107 103% - 111% Tissue Extracts 113 111% - 116% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Bcl-2 ELISA Kit (ab272102) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Bcl-2 protein in human cell and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Bcl-2 with 38.08 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Bcl-2 is the founding member of the apoptotic-regulating Bcl-2 family of proteins. It is considered an apoptosis suppressor in many cell systems and has been shown to interact with many other proteins involved in cell cycle regulation, autophagy, mitochondrial membrane stability, and oxidative stress response, among many others. As such an important protein involved in cell regulation, mutation or damage to the protein’s structure or function are critically linked to a variety of cancers. In particular, a chromosomal translocation occurs in follicular lymphoma which causes the Bcl-2 gene to relocate next to the heavy chain immunoglobulin locus. The unregulated expression of the fusion protein causes an excessively high amount of Bcl-2, disrupting the cell’s apoptosis regulation and leading to unregulated cell growth.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Bcl-2 Capture Antibody 1 x 600µl 10X Human Bcl-2 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human Bcl-2 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. 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). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785). -
Tissue specificity
Expressed in a variety of tissues. -
Involvement in disease
A chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions. -
Sequence similarities
Belongs to the Bcl-2 family. -
Domain
BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.
The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
The loop between motifs BH4 and BH3 is required for the interaction with NLRP1. -
Post-translational
modificationsPhosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
Monoubiquitinated by PARK2, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome. -
Cellular localization
Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane. - Information by UniProt
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Alternative names
- Apoptosis regulator Bcl 2
- Apoptosis regulator Bcl-2
- Apoptosis regulator Bcl2
see all -
Database links
- Entrez Gene: 596 Human
- Omim: 151430 Human
- SwissProt: P10415 Human
- Unigene: 150749 Human
Images
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Human Bcl-2 standard curve comparisonStandard Curve comparison between the original Human Bcl-2 SimpleStep ELISA kit (ab202411) and current Human MMP3 SimpleStep ELISA kit (ab272102).
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Example of human Bcl-2 standard curve in 1X Cell Extraction Buffer PTR.The Bcl-2 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Bcl-2 in human MCF7, U266B1, and thymus homogenate extract samples based on 250, 62.50, and 250 µg/mL extract loads respectively.The concentrations of Bcl-2 were measured in duplicate and interpolated from the Bcl-2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Bcl-2 concentration was determined to be 4,668 pg/mL in MCF7, 7,515 pg/ml in U266B1, and 1,815 pg/ml in thymus homogenate extract.
