GNLY ELISA Kit

Catalog No. ABIN1889422

Human GNLY ELISA Kit Colorimetric assay for quantification of Human GNLY.

Quick Overview for GNLY ELISA Kit (ABIN1889422)

Target: GNLY (Granulysin (GNLY))

Binding Specificity: AA 1-145

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Product Details GNLY ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Granulysin

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: M1-L145

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: M1-L145

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Tissue Specificity: Expressed in natural killer and T-cells.

Plate: Pre-coated

Protocol: human Granulysin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Granulysin has been precoated onto 96-well plates. Standards(NSO, M1-L145) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Granulysin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Granulysin amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL human Granulysin standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human Granulysin standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 355, Standard deviation: 20.59, CV(%): 5.8
• Sample 2: n=16, Mean(pg/ml): 862, Standard deviation: 38.79, CV(%): 4.5
• Sample 3: n=16, Mean(pg/ml): 1513, Standard deviation: 65, CV(%): 4.3,
• Sample 1: n=24, Mean(pg/ml): 324, Standard deviation: 26.5, CV(%): 8.2
• Sample 2: n=24, Mean(pg/ml): 765, Standard deviation: 55.85, CV(%): 7.3
• Sample 3: n=24, Mean(pg/ml): 1322, Standard deviation: 89.8, CV(%): 6.8

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for GNLY

Target: GNLY (Granulysin (GNLY))

Alternative Name: GNLY

Background:

Protein Function: Antimicrobial protein that kills intracellular pathogens. Active against a broad range of microbes, including Gram-positive and Gram-negative bacteria, fungi, and parasites. Kills Mycobacterium tuberculosis.

Background: Granulysin is a substance released by cytotoxic T cells(CD8) when they are attached to infected body cells. The product of this gene is a member of the saposin-like protein(SAPLIP) family. It is mapped to 2p11.2. Granulysin functions to create holes in the target cell membrane and destroy it. It is able to induce apoptosis in target cells and also has antimicrobial action. This gene is expressed in cytolytic granules with perforin, a pore forming protein, and granzymes that are also involved in cytolysis. In addition to it, Granulysin is broadly antimicrobial, killing microbes that cause, for example, tuberculosis and malaria, and can destroy some tumors. A series of peptides generated from the amino acid sequence of Granulysin are potential antibiotics. It has been found that secretory Granulysin is a key molecule responsible for the disseminated keratinocyte death in SJS/TEN.

Synonyms: Granulysin,Lymphokine LAG-2,Protein NKG5,T-cell activation protein 519,GNLY,LAG2, NKG5, TLA519,

Full Gene Name: Granulysin

Cellular Localisation: Secreted. Located in the cytotoxic granules of T-cells, which are released upon antigen stimulation.

Gene ID: 10578

UniProt: P22749