Human GAL3 (Galectin 3) CLIA Kit (HUES00839)

SKU:
HUES00839
Product type:
ELISA
Reactivity:
Human
ELISA Type:
CLIA
€599

Description

ELISA Kit Technical ManualMSDS

Human GAL3 (Galectin 3) 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 GAL3 . 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 GAL3 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 GAL3, 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 GAL3. The concentration of Human GAL3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human GAL3 in samples. No significant cross-reactivity or interference between Human GAL3 and 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
200054460
56982
5572155693
100022317
25631
2397423946
50012024
10040
1103211004
2505126
5462
52945266
1252674
2542
26082580
62.51313
1309
13111283
31.25645
703
674646
027
29
28--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GAL3 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GAL3 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)104.64302.32798.06110.52316.19728.10
Standard deviation11.3025.1572.4612.8830.7074.41
C V (%)10.808.329.0811.659.7110.22

Recovery

The recovery of Human GAL3 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)95-111101
EDTA plasma (n=5)87-10094
Cell culture media (n=5)93-10899

Linearity

Samples were spiked with high concentrations of Human GAL3 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)
1:2Range (%)100-116101-11488-100
Average (%)10610793
1:4Range (%)89-10491-106101-117
Average (%)9698107
1:8Range (%)89-10393-10692-103
Average (%)9410197
1:16Range (%)101-11693-110101-117
Average (%)107100106

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 Standard2 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 Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human GAL3 (Galectin 3) CLIA Kit (HUES00839) 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 GAL3 (Galectin 3) CLIA Kit (HUES00839) Protein Information

UniProt Protein Function:Galectin-3: galactose-specific lectin which binds IgE. Expressed at a high level in the colonic epithelium. Also abundant in activated macrophages.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Extracellular matrix; Cell surface

Chromosomal Location of Human Ortholog: 14q22.3

Cellular Component: spliceosome; extracellular matrix; extracellular space; proteinaceous extracellular matrix; membrane; mitochondrial inner membrane; cytoplasm; plasma membrane; immunological synapse; nucleus; external side of plasma membrane

Molecular Function:protein binding; laminin binding; IgE binding; carbohydrate binding; chemoattractant activity

Biological Process: neutrophil chemotaxis; extracellular matrix organization and biogenesis; RNA splicing; negative regulation of endocytosis; regulation of T cell proliferation; negative regulation of T cell receptor signaling pathway; macrophage chemotaxis; monocyte chemotaxis; epithelial cell differentiation; positive chemotaxis; eosinophil chemotaxis; innate immune response; mRNA processing; skeletal development

NCBI Summary:This gene encodes a member of the galectin family of carbohydrate binding proteins. Members of this protein family have an affinity for beta-galactosides. The encoded protein is characterized by an N-terminal proline-rich tandem repeat domain and a single C-terminal carbohydrate recognition domain. This protein can self-associate through the N-terminal domain allowing it to bind to multivalent saccharide ligands. This protein localizes to the extracellular matrix, the cytoplasm and the nucleus. This protein plays a role in numerous cellular functions including apoptosis, innate immunity, cell adhesion and T-cell regulation. The protein exhibits antimicrobial activity against bacteria and fungi. Alternate splicing results in multiple transcript variants.[provided by RefSeq, Oct 2014]
UniProt Code:P17931
NCBI GenInfo Identifier:215274262
NCBI Gene ID:3958
NCBI Accession:P17931.5
UniProt Secondary Accession:P17931,Q16005, Q6IBA7, Q96J47, B2RC38,
UniProt Related Accession:P17931
Molecular Weight:
NCBI Full Name:Galectin-3
NCBI Synonym Full Names:lectin, galactoside-binding, soluble, 3
NCBI Official Symbol:LGALS3
NCBI Official Synonym Symbols:L31; GAL3; MAC2; CBP35; GALBP; GALIG; LGALS2
NCBI Protein Information:galectin-3; lectin L-29; 35 kDa lectin; MAC-2 antigen; IgE-binding protein; laminin-binding protein; galactose-specific lectin 3; carbohydrate-binding protein 35
UniProt Protein Name:Galectin-3
UniProt Synonym Protein Names:35 kDa lectin; Carbohydrate-binding protein 35; CBP 35; Galactose-specific lectin 3; Galactoside-binding protein; GALBP; IgE-binding protein; L-31; Laminin-binding protein; Lectin L-29; Mac-2 antigen
Protein Family:Galectin
UniProt Gene Name:LGALS3
UniProt Entry Name:LEG3_HUMAN
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