Human/Mouse Total Bcl-xL DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total Bcl-xL in cell lysates. An immobilized capture antibody specific for Bcl-xL binds both phosphorylated and unphosphorylated Bcl-xL. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product
The Human/Mouse Total Bcl-xL DuoSet® IC ELISA is specific for Bcl-xL as shown by Western Blot analysis of the protein bound by the capture antibody supplied in the kit. Lysates prepared from A431 human epithelial carcinoma cells were incubated in wells coated with Total Bcl-xL Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. Captured proteins were electrophoresed, transferred to a PVDF membrane, and immunoblotted with Total Bcl-xL Detection Antibody. Only a single band corresponding to human Bcl-xL was detected.
Amounts of Bcl-xL, as measured by the Human/Mouse Total Bcl-xL DuoSet® IC ELISA, are consistent with the relative amounts of human and mouse Bcl-xL determined by qualitative Western Blot analysis. A431 human epithelial carcinoma cells and NIH-3T3 mouse embryonic fibroblast cells were lysed as described in the Preparation of Samples section in the product insert (Catalog # DYC894). Human and mouse Bcl-xL were measured with this DuoSet® IC ELISA. The same lysates were immunoblotted (inset) with rat anti-mouse Bcl-xL. The DuoSet® IC ELISA results correlate with the relative amounts of Bcl-xL detected by Western Blot.
Preparation and Storage
Bcl-x is a member of the Bcl-2 family of proteins that regulates outer mitochondrial membrane permeability. Alternative splicing results in two distinct Bcl-x isoforms. Bcl-xL (long) is an anti-apoptotic protein that prevents release of Cytochromec from the mitochondrial intermembrane space into the cytosol. Bcl-xs (short) is a pro-apoptotic member that can initiate programmed cell death.
Citations for Human/Mouse Total Bcl-xL DuoSet IC ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 4
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Bacteria differentially induce degradation of Bcl-xL, a survival protein, by human platelets.
Authors: Kraemer B, Campbell R, Schwertz H, Franks Z, Vieira de Abreu A, Grundler K, Kile B, Dhakal B, Rondina M, Kahr W, Mulvey M, Blaylock R, Zimmerman G, Weyrich A
Sample Types: Whole Cells
Engagement of CD31 delivers an activating signal that contributes to the survival of chronic lymphocytic leukaemia cells.
Authors: Poggi A, Prevosto C, Catellani S, Rocco I, Garuti A, Zocchi MR
Br. J. Haematol., 2010;151(3):252-64.
Sample Types: Cell Lysates
Systemic delivery of siRNA specific to tumor mediated by atelocollagen: combined therapy using siRNA targeting Bcl-xL and cisplatin against prostate cancer.
Authors: Mu P, Nagahara S, Makita N, Tarumi Y, Kadomatsu K, Takei Y
Int. J. Cancer, 2009;125(12):2978-90.
Sample Types: Tissue Homogenates
AAV-mediated expression of Bcl-xL or XIAP fails to induce neuronal resistance against quinolinic acid-induced striatal lesioning.
Authors: Kells AP, Connor B
Neurosci. Lett., 2008;436(3):326-30.
Sample Types: Tissue Homogenates
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