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ELISAGenie/Human TGM3 (Transglutaminase 3, Epidermal) CLIA Kit (HUES01210)/Transglutaminase 3, Epidermal) CLIA Kit (HUES01210/HUES01210
ELISAGenie/Human TGM3 (Transglutaminase 3, Epidermal) CLIA Kit (HUES01210)/Transglutaminase 3, Epidermal) CLIA Kit (HUES01210/HUES01210
- 产品编号： HUES01210
- 美 元 价： $599.00
- 市 场 价： ¥11980.00
- 品 牌： ELISAGenie
- 产 地： 美国(厂家直采)
- 公 司： ELISAGenie
- 产品分类： 试剂盒>纯化试剂盒>病毒纯化试剂盒>
- 公司分类：
- 联系 Q Q: 1570468124
- 电话号码: 4000-520-616
- 邮 箱: info@ebiomall.com
- Description
- Additional Information
Description
Human TGM3 (Transglutaminase 3, Epidermal) CLIA Kit
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human TGM3 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TGM3 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 TGM3, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human TGM3. The concentration of Human TGM3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| Assay type | Sandwich |
| Format | 96T |
| Assay time | 4.5h |
| Reactivity | Human |
| Detection method | Chemiluminescence |
| Detection range | 62.50-4000 pg/mL |
| Sensitivity | 37.50 pg/mL |
| Sample volume | 100uL |
| Sample type | Serum, plasma and other biological fluids |
| Repeatability | CV < 15% |
Specificity
This kit recognizes Human TGM3in samples. No significant cross-reactivity or interference between Human TGM3and analogues was observed.
Typical Data
As the RLU 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(pg/mL) | RLU | Average | Corrected |
|---|---|---|---|
| 4000 | 5762660536 | 59081 | 59046 |
| 2000 | 2703528683 | 27859 | 27824 |
| 1000 | 1449412474 | 13484 | 13449 |
| 500 | 61787032 | 6605 | 6570 |
| 250 | 33413145 | 3243 | 3208 |
| 125 | 17051457 | 1581 | 1546 |
| 62.50 | 748762 | 755 | 720 |
| 0 | 3337 | 35 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TGM3were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TGM3were 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 (pg/mL) | 181.87 | 340.40 | 1927.05 | 177.18 | 349.85 | 2090.54 |
| Standard deviation | 15.24 | 32.51 | 123.91 | 14.39 | 27.15 | 130.24 |
| C V (%) | 8.38 | 9.55 | 6.43 | 8.12 | 7.76 | 6.23 |
Recovery
The recovery of Human TGM3spiked 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) | 87-101 | 92 |
| EDTA plasma (n=5) | 99-117 | 107 |
| Cell culture media (n=5) | 85-97 | 91 |
Linearity
Samples were spiked with high concentrations of Human TGM3and 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 (%) | 101-119 | 88-100 | 100-119 |
| Average (%) | 109 | 93 | 108 | |
| 1:4 | Range (%) | 85-100 | 87-103 | 99-115 |
| Average (%) | 92 | 95 | 106 | |
| 1:8 | Range (%) | 95-109 | 101-113 | 94-110 |
| Average (%) | 100 | 107 | 102 | |
| 1:16 | Range (%) | 86-100 | 87-98 | 85-101 |
| Average (%) | 93 | 92 | 92 |
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 CLIA Plate(Dismountable) | 8 wells X12 strips | -20"C, 6 months |
| Reference Standard | 2 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 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 (25X) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4"C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4"C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
Human TGM3 (Transglutaminase 3, Epidermal) CLIA Kit (HUES01210) 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 100µl of standard solutions into the standard wells.
- 3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- 4. Add 100µl 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 100µL 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 350µL 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 100µL 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- 11. Determine the RLU value of each well immediately.
Human TGM3 (Transglutaminase 3, Epidermal) CLIA Kit (HUES01210) Protein Information
| UniProt Protein Function: | TGM3: Catalyzes the calcium-dependent formation of isopeptide cross-links between glutamine and lysine residues in various proteins, as well as the conjugation of polyamines to proteins. Involved in the formation of the cornified envelope (CE), a specialized component consisting of covalent cross-links of proteins beneath the plasma membrane of terminally differentiated keratinocytes. Catalyzes small proline-rich proteins (SPRR1 and SPRR2) and LOR cross-linking to form small interchain oligomers, which are further cross-linked by TGM1 onto the growing CE scaffold. In hair follicles, involved in cross- linking structural proteins to hardening the inner root sheath. Belongs to the transglutaminase superfamily. Transglutaminase family. |
| UniProt Protein Details: | Protein type:EC 2.3.2.13; Membrane protein, peripheral; Transferase Chromosomal Location of Human Ortholog: 20q11.2 Cellular Component: extrinsic to internal side of plasma membrane; cytoplasm Molecular Function:protein-glutamine gamma-glutamyltransferase activity; transferase activity, transferring acyl groups; calcium ion binding; catalytic activity Biological Process: keratinocyte differentiation; cell envelope organization and biogenesis; hair follicle morphogenesis; keratinization; protein modification process; peptide cross-linking; protein tetramerization |
| NCBI Summary: | Transglutaminases are enzymes that catalyze the crosslinking of proteins by epsilon-gamma glutamyl lysine isopeptide bonds. While the primary structure of transglutaminases is not conserved, they all have the same amino acid sequence at their active sites and their activity is calcium-dependent. The protein encoded by this gene consists of two polypeptide chains activated from a single precursor protein by proteolysis. The encoded protein is involved the later stages of cell envelope formation in the epidermis and hair follicle. [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q08188 |
| NCBI GenInfo Identifier: | 257051080 |
| NCBI Gene ID: | 7053 |
| NCBI Accession: | Q08188.4 |
| UniProt Secondary Accession: | Q08188,O95933, Q32ML9, Q32MM0, A8K5N6, B2RCR6, D3DVX1 |
| UniProt Related Accession: | Q08188 |
| Molecular Weight: | |
| NCBI Full Name: | Protein-glutamine gamma-glutamyltransferase E |
| NCBI Synonym Full Names: | transglutaminase 3 |
| NCBI Official Symbol: | TGM3 |
| NCBI Official Synonym Symbols: | TGE |
| NCBI Protein Information: | protein-glutamine gamma-glutamyltransferase E; TG(E); TGase E; TGase-3; transglutaminase E; transglutaminase-3; E polypeptide, protein-glutamine-gamma-glutamyltransferase |
| UniProt Protein Name: | Protein-glutamine gamma-glutamyltransferase E |
| UniProt Synonym Protein Names: | Transglutaminase E; TG(E); TGE; TGase E; Transglutaminase-3; TGase-3 |
| Protein Family: | Protein-glutamine gamma-glutamyltransferase |
| UniProt Gene Name: | TGM3 |
| UniProt Entry Name: | TGM3_HUMAN |
Additional Information
Reactivity: | Human |
ELISA Type: | CLIA |
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