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Human Caspase-14 ELISA Kit

SKU:
HUFI01860
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P31944
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
CASP14, Caspase-14, CASP-14, CASP14, apoptosis-related cysteine protease, caspase 14, apoptosis-related cysteine peptidase, caspase 14, apoptosis-related cysteine protease, caspase-14, MICE
Reactivity:
Human
Research Area:
Developmental Biology
€599
Frequently bought together:

Description

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Human Caspase-14 ELISA Kit

The Human Caspase-14 ELISA Kit is a highly reliable tool for the precise measurement of caspase-14 levels in human samples including serum, plasma, and cell culture supernatants. With its exceptional sensitivity and specificity, this kit delivers accurate and consistent results, making it an invaluable resource for various research applications.Caspase-14 is a key player in programmed cell death, or apoptosis, and is involved in regulating skin barrier function and inflammation.

Dysregulation of caspase-14 has been linked to skin disorders such as psoriasis and atopic dermatitis, highlighting its importance as a potential therapeutic target and diagnostic marker.By utilizing the Human Caspase-14 ELISA Kit, researchers can gain valuable insights into the role of caspase-14 in skin health and disease, paving the way for the development of novel treatments and interventions. Stay ahead in your research with this advanced ELISA kit from Assay Genie.

Product Name:Human Caspase-14 ELISA Kit
Product Code:HUFI01860
Size:96 Assays
Alias:CASP14, Caspase-14, CASP-14, CASP14, apoptosis-related cysteine protease, caspase 14, apoptosis-related cysteine peptidase, caspase 14, apoptosis-related cysteine protease, caspase-14, MICE
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CASP14 concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CASP14 and the recovery rates were calculated by comparing the measured value to the expected amount of Human CASP14 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)94-10197
EDTA plasma(n=5)88-10495
UFH plasma(n=5)86-9893
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CASP14 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-104%87-101%85-103%
EDTA plasma(n=5)83-97%88-99%82-89%
UFH plasma(n=5)84-98%82-88%85-99%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP31944
UniProt Protein Function:CASP14: Believed to be a non-apoptotic caspase which is involved in epidermal differentiation. Seems to play a role in keratinocyte differentiation and cornification. Probably regulates maturation of the epidermis by proteolytically processing filaggrin. Complex of unprocessed caspase-14 and processed 19 kDa (p19) and 10 kDa (p10) subunits. In undifferentiated keratinocytes under postconfluency growth conditions (in vitro). Expressed in keratinocytes of adult skin suprabasal layers (from spinous layers to the stratum granulosum and stratum corneum). Expressed in keratinocytes of hair shaft and sebaceous glands. In psoriatic skin only expressed at very low levels. Belongs to the peptidase C14A family.
UniProt Protein Details:Protein type:EC 3.4.22.-; Protease

Chromosomal Location of Human Ortholog: 19p13.1

Cellular Component: cytoplasm; keratin filament; nucleus

Molecular Function:cysteine-type endopeptidase activity

Biological Process: epidermis development; DNA damage response, signal transduction resulting in induction of apoptosis; keratinization; proteolysis

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. This caspase has been shown to be processed and activated by caspase 8 and caspase 10 in vitro, and by anti-Fas agonist antibody or TNF-related apoptosis inducing ligand in vivo. The expression and processing of this caspase may be involved in keratinocyte terminal differentiation, which is important for the formation of the skin barrier. [provided by RefSeq, Jul 2008]
UniProt Code:P31944
NCBI GenInfo Identifier:12231007
NCBI Gene ID:23581
NCBI Accession:P31944.2
UniProt Secondary Accession:P31944,O95823, Q3SYC9,
UniProt Related Accession:P31944
Molecular Weight:27,680 Da
NCBI Full Name:Caspase-14
NCBI Synonym Full Names:caspase 14, apoptosis-related cysteine peptidase
NCBI Official Symbol:CASP14
NCBI Protein Information:caspase-14; CASP-14; apoptosis-related cysteine protease; caspase 14, apoptosis-related cysteine protease
UniProt Protein Name:Caspase-14
Protein Family:Caspase
UniProt Gene Name:CASP14
UniProt Entry Name:CASPE_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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