Description
Human Bcl-10 (B-cell Leukemia/Lymphoma 10) ELISA Kit
The Human BCL-10 (B-Cell Leukemia/Lymphoma 10) ELISA Kit is specifically designed for the accurate measurement of BCL-10 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and dependable results for a variety of research applications.BCL-10 is a critical protein involved in regulating immune responses and cell survival, particularly in B-cell lymphomas and leukemias.
Its dysregulation is linked to various autoimmune diseases and cancers, underscoring its importance as a biomarker for disease diagnosis and therapeutic development.With this ELISA kit, researchers can confidently study the role of BCL-10 in disease pathogenesis and progression, leading to potential advancements in treatment strategies. Trust in the reliability of the Human BCL-10 ELISA Kit for your research needs.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 1.56-100 ng/mL |
| Sensitivity: | 0.94 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human Bcl-10 in samples. No significant cross-reactivity or interference between Human Bcl-10 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 Human Bcl-10. 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 Bcl-10 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 Bcl-10, 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 Bcl-10. The concentration of Human Bcl-10 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | Bcl-10: a ubiquitous pro-apoptotic protein that promotes pro-caspase-9 maturation and activation of NF-kappa-B via NIK and IKK. May be an adapter protein between upstream TNFR1-TRADD-RIP complex and the downstream NIK-IKK-IKAP complex. Self-associates via CARD-CARD interactions; forms a tight complex with MALT1. Interacts with other CARD-proteins such as CARD9, CARD10, CARD11 and CARD14. Binds caspase-9 with its C-terminal domain. Interacts with TRAF2 and BIRC2/c-IAP2. Defects in BCL10 are involved in various types of cancer. |
| UniProt Protein Details: | Protein type:Apoptosis; Tumor suppressor; Transcription, coactivator/corepressor Chromosomal Location of Human Ortholog: 1p22 Cellular Component: CBM complex; protein complex; lysosome; immunological synapse; cytosol; lipid raft; T cell receptor complex; cytoplasmic microtubule; perinuclear region of cytoplasm; cytoplasm; plasma membrane; lipopolysaccharide receptor complex; nucleus Molecular Function:protein C-terminus binding; protein kinase B binding; ubiquitin binding; protease binding; protein self-association; transcription coactivator activity; transcription factor binding; protein kinase binding; NF-kappaB binding; protein binding; enzyme binding; ubiquitin protein ligase binding; kinase activator activity; kinase binding Biological Process: cell death; response to fungus; positive regulation of transcription, DNA-dependent; positive regulation of caspase activity; B cell apoptosis; T cell receptor signaling pathway; interleukin-6 biosynthetic process; activation of NF-kappaB transcription factor; negative regulation of mature B cell apoptosis; response to molecule of bacterial origin; protein homooligomerization; response to food; adaptive immune response; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-8 biosynthetic process; protein oligomerization; positive regulation of protein ubiquitination; lymphotoxin A biosynthetic process; regulation of T cell receptor signaling pathway; neural tube closure; toll-like receptor signaling pathway; innate immune response; cellular defense response; positive regulation of mast cell cytokine production; positive regulation of T cell activation; immunoglobulin mediated immune response; positive regulation of phosphorylation Disease: Follicular Lymphoma, Susceptibility To, 1; Mesothelioma, Malignant; Gastric Lymphoma, Primary; Testicular Germ Cell Tumor; Immunodeficiency 37 |
| NCBI Summary: | This gene was identified by its translocation in a case of mucosa-associated lymphoid tissue (MALT) lymphoma. The protein encoded by this gene contains a caspase recruitment domain (CARD), and has been shown to induce apoptosis and to activate NF-kappaB. This protein is reported to interact with other CARD domain containing proteins including CARD9, 10, 11 and 14, which are thought to function as upstream regulators in NF-kappaB signaling. This protein is found to form a complex with MALT1, a protein encoded by another gene known to be translocated in MALT lymphoma. MALT1 and this protein are thought to synergize in the activation of NF-kappaB, and the deregulation of either of them may contribute to the same pathogenetic process that leads to the malignancy. [provided by RefSeq, Jul 2008] |
| UniProt Code: | O95999 |
| NCBI GenInfo Identifier: | 4502379 |
| NCBI Gene ID: | 8915 |
| NCBI Accession: | NP_003912. 1 |
| UniProt Secondary Accession: | O95999,Q5VUF1, |
| UniProt Related Accession: | O95999 |
| Molecular Weight: | 26,252 Da |
| NCBI Full Name: | B-cell lymphoma/leukemia 10 |
| NCBI Synonym Full Names: | B-cell CLL/lymphoma 10 |
| NCBI Official Symbol: | BCL10 |
| NCBI Official Synonym Symbols: | CLAP; mE10; CIPER; c-E10; CARMEN |
| NCBI Protein Information: | B-cell lymphoma/leukemia 10; hCLAP; cCARMEN; cellular-E10; cellular homolog of vCARMEN; CARD-containing proapoptotic protein; CARD containing molecule enhancing NF-kB; CARD-containing apoptotic signaling protein; caspase-recruiting domain-containing prote |
| UniProt Protein Name: | B-cell lymphoma/leukemia 10 |
| UniProt Synonym Protein Names: | B-cell CLL/lymphoma 10; Bcl-10; CARD-containing molecule enhancing NF-kappa-B; CARD-like apoptotic protein; hCLAP; CED-3/ICH-1 prodomain homologous E10-like regulator; CIPER; Cellular homolog of vCARMEN; cCARMEN; Cellular-E10; c-E10; Mammalian CARD-containing adapter molecule E10 |
| Protein Family: | B-cell lymphoma/leukemia |
| UniProt Gene Name: | BCL10 |
| UniProt Entry Name: | BCL10_HUMAN |
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 |
| 100 | 2.468 2.496 | 2.482 | 2.418 |
| 50 | 1.636 1.648 | 1.642 | 1.578 |
| 25 | 0.964 0.964 | 0.964 | 0.9 |
| 12.5 | 0.505 0.523 | 0.514 | 0.45 |
| 6.25 | 0.258 0.252 | 0.255 | 0.191 |
| 3.13 | 0.176 0.152 | 0.164 | 0.1 |
| 1.56 | 0.113 0.117 | 0.115 | 0.051 |
| 0 | 0.056 0.072 | 0.064 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human Bcl-10 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Bcl-10 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 (ng/mL) | 4.98 | 14.83 | 43.26 | 4.97 | 15.75 | 42.87 |
| Standard deviation | 0.29 | 0.65 | 1.72 | 0.31 | 0.83 | 2.27 |
| C V (%) | 5.82 | 4.38 | 3.98 | 6.24 | 5.27 | 5.30 |
Recovery
The recovery of Human Bcl-10 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) | 96-108 | 101 |
| EDTA plasma (n=5) | 92-109 | 99 |
| Cell culture media (n=5) | 91-105 | 96 |
Linearity
Samples were spiked with high concentrations of Human Bcl-10 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 (%) | 95-111 | 84-96 | 93-104 |
| Average (%) | 103 | 90 | 99 | |
| 1:4 | Range (%) | 94-104 | 83-95 | 83-93 |
| Average (%) | 99 | 90 | 88 | |
| 1:8 | Range (%) | 92-105 | 85-99 | 84-95 |
| Average (%) | 99 | 92 | 90 | |
| 1:16 | Range (%) | 86-100 | 81-91 | 87-98 |
| Average (%) | 92 | 85 | 92 |
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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 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.
- 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.
- Add 100 µL 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 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.
- 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.
