Human CASP7 (Caspase 7) ELISA Kit (HUES01820)
- SKU:
- HUES01820
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P55210
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Death
Description
Human CASP7 (Caspase 7) ELISA Kit
The Human Caspase-7 (CASP7) ELISA Kit is specifically designed for the accurate measurement of caspase-7 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for various research applications.Caspase-7 is a key enzyme involved in the process of apoptosis, playing a critical role in programmed cell death. Dysregulation of caspase-7 has been linked to various diseases, including cancer, neurodegenerative disorders, and autoimmune conditions.
As such, measuring caspase-7 levels can provide valuable insights for understanding disease mechanisms and developing potential therapeutic interventions.Overall, the Human Caspase-7 (CASP7) ELISA Kit is a valuable tool for researchers seeking to explore the role of caspase-7 in health and disease, offering reliable and accurate measurements to support their studies.
Assay type: | Sandwich |
Format: | 96T |
Assay time: | 4.5h |
Reactivity: | Human |
Detection Method: | Colormetric |
Detection Range: | 0.31-20 ng/mL |
Sensitivity: | 0.19 ng/mL |
Sample Volume Required Per Well: | 100µL |
Sample Type: | Serum, plasma and other biological fluids |
Specificity: | This kit recognizes Human CASP7 in samples. No significant cross-reactivity or interference between Human CASP7 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 CASP7. 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 CASP7 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 CASP7, 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 CASP7. The concentration of Human CASP7 in samples can be calculated by comparing the OD of the samples to the standard curve.
UniProt Protein Function: | CASP7: Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly- 217' bond. Overexpression promotes programmed cell death. Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 20 kDa (p20) and a 11 kDa (p11) subunit. Interacts with BIRC6/bruce. Highly expressed in lung, skeletal muscle, liver, kidney, spleen and heart, and moderately in testis. No expression in the brain. Inhibited by isatin sulfonamides. Belongs to the peptidase C14A family. 3 isoforms of the human protein are produced by alternative splicing. |
UniProt Protein Details: | Protein type:Apoptosis; EC 3. 4. 22. 60; Endoplasmic reticulum; Protease; Mitochondrial Chromosomal Location of Human Ortholog: 10q25 Cellular Component: nucleoplasm; cytoplasm; nucleus; cytosol Molecular Function:protein binding; cysteine-type endopeptidase activity; cysteine-type peptidase activity Biological Process: apoptosis; caspase activation via cytochrome c; proteolysis; cell structure disassembly during apoptosis |
NCBI Summary: | This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. The precursor of the encoded protein is cleaved by caspase 3 and 10, is activated upon cell death stimuli and induces apoptosis. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, May 2012] |
UniProt Code: | P55210 |
NCBI GenInfo Identifier: | 1730092 |
NCBI Gene ID: | 840 |
NCBI Accession: | P55210. 1 |
UniProt Secondary Accession: | P55210,Q13364, Q53YD5, Q5SVL0, Q5SVL3, Q96BA0, B4DQU7 B5BU45, D3DRB8, |
UniProt Related Accession: | P55210 |
Molecular Weight: | 31,614 Da |
NCBI Full Name: | Caspase-7 |
NCBI Synonym Full Names: | caspase 7, apoptosis-related cysteine peptidase |
NCBI Official Symbol: | CASP7 |
NCBI Official Synonym Symbols: | MCH3; CMH-1; LICE2; CASP-7; ICE-LAP3 |
NCBI Protein Information: | caspase-7; apoptotic protease MCH-3; ICE-like apoptotic protease 3; caspase 7, apoptosis-related cysteine protease |
UniProt Protein Name: | Caspase-7 |
UniProt Synonym Protein Names: | Apoptotic protease Mch-3; CMH-1; ICE-like apoptotic protease 3; ICE-LAP3 |
Protein Family: | Caspase |
UniProt Gene Name: | CASP7 |
UniProt Entry Name: | CASP7_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 |
20 | 2.514 2.534 | 2.524 | 2.464 |
10 | 1.599 1.649 | 1.624 | 1.564 |
5 | 0.987 0.985 | 0.986 | 0.926 |
2.5 | 0.46 0.496 | 0.478 | 0.418 |
1.25 | 0.261 0.247 | 0.254 | 0.194 |
0.63 | 0.169 0.159 | 0.164 | 0.104 |
0.31 | 0.113 0.113 | 0.113 | 0.053 |
0 | 0.055 0.065 | 0.06 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CASP7 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CASP7 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) | 1.01 | 2.88 | 8.96 | 0.93 | 2.73 | 8.43 |
Standard deviation | 0.07 | 0.13 | 0.36 | 0.06 | 0.12 | 0.41 |
C V (%) | 6.93 | 4.51 | 4.02 | 6.45 | 4.40 | 4.86 |
Recovery
The recovery of Human CASP7 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) | 88-102 | 96 |
EDTA plasma (n=5) | 87-99 | 93 |
Cell culture media (n=5) | 88-99 | 93 |
Linearity
Samples were spiked with high concentrations of Human CASP7 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 (%) | 89-104 | 99-112 | 94-106 |
Average (%) | 95 | 105 | 99 | |
1:4 | Range (%) | 93-109 | 85-99 | 89-101 |
Average (%) | 100 | 90 | 94 | |
1:8 | Range (%) | 87-100 | 80-94 | 84-97 |
Average (%) | 93 | 86 | 90 | |
1:16 | Range (%) | 91-105 | 80-94 | 85-96 |
Average (%) | 97 | 86 | 89 |
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.