Human/Mouse Total Bcl-xL DuoSet IC ELISA

R&D Systems | Catalog # DYC894-2

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

625-20000 pg/mL

Sample Type

Cell Lysates

Reactivity

Human, Mouse

Human/Mouse Total Bcl-xL DuoSet IC ELISA Features

  • 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
View all Bcl-xL ELISA Kits »

Product Summary for Human/Mouse Total Bcl-xL DuoSet IC ELISA

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.

 

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

Cell lysates (100 µL)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human/Mouse Total Bcl-xL DuoSet IC ELISA

Standard Curve

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.

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.

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.

Kit Contents for Human/Mouse Total Bcl-xL DuoSet IC ELISA

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

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

Lysis Buffer*

IC Diluent*

Blocking Buffer*

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

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Bcl-xL

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.

Long Name

B Cell Lymphoma/Leukemia x Long Form

Alternate Names

BclxL

Entrez Gene IDs

598 (Human); 12048 (Mouse); 24888 (Rat)

Gene Symbol

BCL2L1

Additional Bcl-xL Products

Product Documents for Human/Mouse Total Bcl-xL DuoSet IC ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse Total Bcl-xL DuoSet IC ELISA

For research use only

Citations for Human/Mouse Total Bcl-xL DuoSet IC ELISA

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