Human CRT (Calreticulin) ELISA Kit (HUES01790)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- Calreticulin, calregulin,CRP55,CaBP3,calsequestrin-like protein, endoplasmic reticulum resident protein 60 ,ERp60,CALR, CRT, RO, SSA, cC1qR
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.16-10 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human CRT in samples. No significant cross-reactivity or interference between Human CRT 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 CRT. 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 CRT 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 CRT, 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 CRT. The concentration of Human CRT in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||Calreticulin: Calcium-binding chaperone that promotes folding, oligomeric assembly and quality control in the endoplasmic reticulum (ER) via the calreticulin/calnexin cycle. This lectin interacts transiently with almost all of the monoglucosylated glycoproteins that are synthesized in the ER. Interacts with the DNA-binding domain of NR3C1 and mediates its nuclear export. Involved in maternal gene expression regulation. May participate in oocyte maturation via the regulation of calcium homeostasis. Monomer. Component of an EIF2 complex at least composed of CELF1/CUGBP1, CALR, CALR3, EIF2S1, EIF2S2, HSP90B1 and HSPA5. Interacts with PDIA3/ERp57. Interacts with NR3C1 and TRIM21. Interacts with GABARAP. Belongs to the calreticulin family.|
|UniProt Protein Details:|
Protein type:Calcium-binding; Motility/polarity/chemotaxis; Nuclear receptor co-regulator; Secreted; Secreted, signal peptide
Chromosomal Location of Human Ortholog: 19p13. 13
Cellular Component: cell surface; cytoplasm; cytosol; endoplasmic reticulum; endoplasmic reticulum lumen; extracellular region; extracellular space; focal adhesion; intracellular; membrane; nucleus; perinuclear region of cytoplasm; polysome
Molecular Function:androgen receptor binding; calcium ion binding; carbohydrate binding; chaperone binding; complement component C1q binding; glycoprotein binding; integrin binding; mRNA binding; protein binding; ubiquitin protein ligase binding; unfolded protein binding; zinc ion binding
Biological Process: antigen processing and presentation of peptide antigen via MHC class I; cellular calcium ion homeostasis; glucocorticoid receptor signaling pathway; negative regulation of neuron differentiation; negative regulation of retinoic acid receptor signaling pathway; negative regulation of steroid hormone receptor signaling pathway; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; negative regulation of translation; peptide antigen assembly with MHC class I protein complex; positive regulation of cell cycle; positive regulation of cell proliferation; positive regulation of DNA replication; positive regulation of phagocytosis; protein export from nucleus; protein folding; protein maturation via protein folding; protein stabilization; receptor-mediated endocytosis; regulation of apoptosis; regulation of transcription, DNA-dependent; sequestering of calcium ion
Disease: Myelofibrosis; Thrombocythemia 1
|NCBI Summary:||Calreticulin is a multifunctional protein that acts as a major Ca(2+)-binding (storage) protein in the lumen of the endoplasmic reticulum. It is also found in the nucleus, suggesting that it may have a role in transcription regulation. Calreticulin binds to the synthetic peptide KLGFFKR, which is almost identical to an amino acid sequence in the DNA-binding domain of the superfamily of nuclear receptors. Calreticulin binds to antibodies in certain sera of systemic lupus and Sjogren patients which contain anti-Ro/SSA antibodies, it is highly conserved among species, and it is located in the endoplasmic and sarcoplasmic reticulum where it may bind calcium. The amino terminus of calreticulin interacts with the DNA-binding domain of the glucocorticoid receptor and prevents the receptor from binding to its specific glucocorticoid response element. Calreticulin can inhibit the binding of androgen receptor to its hormone-responsive DNA element and can inhibit androgen receptor and retinoic acid receptor transcriptional activities in vivo, as well as retinoic acid-induced neuronal differentiation. Thus, calreticulin can act as an important modulator of the regulation of gene transcription by nuclear hormone receptors. Systemic lupus erythematosus is associated with increased autoantibody titers against calreticulin but calreticulin is not a Ro/SS-A antigen. Earlier papers referred to calreticulin as an Ro/SS-A antigen but this was later disproven. Increased autoantibody titer against human calreticulin is found in infants with complete congenital heart block of both the IgG and IgM classes. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||117501|
|NCBI Gene ID:||811|
|NCBI Accession:||P27797. 1|
|UniProt Secondary Accession:||P27797,Q6IAT4, Q9UDG2,|
|UniProt Related Accession:||P27797|
|Molecular Weight:||48,142 Da|
|NCBI Full Name:||Calreticulin|
|NCBI Synonym Full Names:||calreticulin|
|NCBI Official Symbol:||CALR|
|NCBI Official Synonym Symbols:||RO; CRT; SSA; cC1qR; HEL-S-99n|
|NCBI Protein Information:||calreticulin|
|UniProt Protein Name:||Calreticulin|
|UniProt Synonym Protein Names:||CRP55; Calregulin; Endoplasmic reticulum resident protein 60; ERp60; HACBP; grp60|
|UniProt Gene Name:||CALR|
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CRT were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CRT were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.66||4.41||5.26||5.77||4.58||3.42|
The recovery of Human CRT spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||86-98||91|
|Cell culture media (n=5)||90-105||96|
Samples were spiked with high concentrations of Human CRT 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)|
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.
|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.