Human Bcl2L11 (Bcl-2 Like Protein 11) ELISA Kit (HUES01757)

SKU:
HUES01757
€539

Description

ELISA Kit Technical ManualMSDS

Human Bcl2L11 (Bcl-2 Like Protein 11) ELISA Kit

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 Human Bcl2L11 . 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 Human Bcl2L11 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 Human Bcl2L11, 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 Human Bcl2L11. The concentration of Human Bcl2L11 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range0.31-20 ng/mL
Sensitivity0.19 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human Bcl2L11 in samples. No significant cross-reactivity or interference between Human Bcl2L11 and analogues was observed.

Typical Data

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
(ng/mL)
O.D Average Corrected
202.287
2.321
2.3042.249
101.613
1.617
1.6151.56
50.909
0.901
0.9050.85
2.50.414
0.416
0.4150.36
1.250.216
0.208
0.2120.157
0.630.164
0.144
0.1540.099
0.310.102
0.108
0.1050.05
00.054
0.056
0.055--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human Bcl2L11 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Bcl2L11 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)1.082.998.491.033.177.89
Standard deviation0.050.170.360.060.170.33
C V (%)4.635.694.245.835.364.18

Recovery

The recovery of Human Bcl2L11 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)87-10092
EDTA plasma (n=5)84-9590
Cell culture media (n=5)94-108100

Linearity

Samples were spiked with high concentrations of Human Bcl2L11 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)
1:2Range (%)84-9695-10895-110
Average (%)91101102
1:4Range (%)86-9985-10082-94
Average (%)939188
1:8Range (%)90-10380-9385-97
Average (%)978690
1:16Range (%)86-9784-9786-101
Average (%)929193

Kit Components & Storage

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 X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human Bcl2L11 (Bcl-2 Like Protein 11) ELISA Kit (HUES01757) Assay procedure

    1. 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.
    2. Aliquot 100 µL of standard solutions into the standard wells.
    3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL 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.
    7. Aspirate or decant the solution from the plate and add 350 µL 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.
    8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90 µL 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.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human Bcl2L11 (Bcl-2 Like Protein 11) ELISA Kit (HUES01757) Protein Information

UniProt Protein Function:BIM: a pro-apoptotic member of the BCL-2 protein family. Interacts with other members of the BCL-2 protein family, including BCL2, BCL2L1/BCL-X(L), and MCL1, and act as an apoptotic activator. Its expression can be induced by nerve growth factor (NGF), as well as by the forkhead transcription factor FKHR-L1, which suggests a role in neuronal and lymphocyte apoptosis. Transgenic studies in the mouse suggested that this protein functions as an essential initiator of the apoptosis in thymocyte-negative selection. Nineteen alternatively spliced transcript variants of this gene have been reported.
UniProt Protein Details:

Protein type:Apoptosis

Chromosomal Location of Human Ortholog: 2q13

Cellular Component: microtubule; mitochondrial outer membrane; extrinsic to membrane; endomembrane system; cytosol

Molecular Function:protein binding; microtubule binding

Biological Process: nerve growth factor receptor signaling pathway; positive regulation of apoptosis; apoptosis; cell-matrix adhesion; pigmentation during development; myeloid cell homeostasis; ear development; positive regulation of caspase activity; B cell apoptosis; cellular process regulating host cell cycle in response to virus; positive regulation of apoptosis by virus; mammary gland development; B cell homeostasis; positive regulation of neuron apoptosis; T cell homeostasis; kidney development; post-embryonic organ morphogenesis; caspase activation; spleen development; thymus development; in utero embryonic development; positive regulation of protein homooligomerization; male gonad development; positive regulation of cell cycle; regulation of organ growth; lumen formation; odontogenesis of dentine-containing teeth; DNA damage response, signal transduction resulting in induction of apoptosis; regulation of pigmentation during development; spermatogenesis; brain development

NCBI Summary:The protein encoded by this gene belongs to the BCL-2 protein family. BCL-2 family members form hetero- or homodimers and act as anti- or pro-apoptotic regulators that are involved in a wide variety of cellular activities. The protein encoded by this gene contains a Bcl-2 homology domain 3 (BH3). It has been shown to interact with other members of the BCL-2 protein family and to act as an apoptotic activator. The expression of this gene can be induced by nerve growth factor (NGF), as well as by the forkhead transcription factor FKHR-L1, which suggests a role of this gene in neuronal and lymphocyte apoptosis. Transgenic studies of the mouse counterpart suggested that this gene functions as an essential initiator of apoptosis in thymocyte-negative selection. Several alternatively spliced transcript variants of this gene have been identified. [provided by RefSeq, Jun 2013]
UniProt Code:O43521
NCBI GenInfo Identifier:20336315
NCBI Gene ID:10018
NCBI Accession:NP_619527.1
UniProt Secondary Accession:O43521,O43522, Q0MSE7, Q0MSE8, Q0MSE9, Q53R28, Q6JTU6 Q6T851, Q6TE14, Q6TE15, Q6TE16, A8K2W2,
UniProt Related Accession:O43521
Molecular Weight:22,171 Da
NCBI Full Name:bcl-2-like protein 11 isoform 1
NCBI Synonym Full Names:BCL2-like 11 (apoptosis facilitator)
NCBI Official Symbol:BCL2L11
NCBI Official Synonym Symbols:BAM; BIM; BOD
NCBI Protein Information:bcl-2-like protein 11; bcl-2 interacting protein Bim; bcl-2-related ovarian death agonist; bcl-2 interacting mediator of cell death
UniProt Protein Name:Bcl-2-like protein 11
UniProt Synonym Protein Names:Bcl2-interacting mediator of cell death
Protein Family:Bcl-2-like protein
UniProt Gene Name:BCL2L11
UniProt Entry Name:B2L11_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
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