Lipocalin 2 ELISA Kit

Catalog No. ABIN411401

Product Details Lipocalin 2 ELISA Kit

Target: Lipocalin 2 (LCN2)

Binding Specificity: AA 21-198

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10000 pg/mL

Minimum Detection Limit: 156 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Lipocalin-2/NGAL

Brand: PicoKine™

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

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: Q21-G198

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: Q21-G198

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:

Sequence similarities: Belongs to the calycin superfamily. Lipocalin family.

Tissue Specificity: Expressed in bone marrow and in tissues that are prone to exposure to microorganism. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. Not found in the small intestine or peripheral blood leukocytes. .

Plate: Pre-coated

Protocol: human NGAL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for NGAL has been precoated onto 96-well plates. Standards(NSO, Q21-G198) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for NGAL 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 NGAL amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 10000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL human NGAL 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, plasma(heparin), saliva or urine to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human NGAL standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 1246, Standard deviation: 48.6, CV(%): 3.9
• Sample 2: n=16, Mean(pg/ml): 3486, Standard deviation: 174.3, CV(%): 5
• Sample 3: n=16, Mean(pg/ml): 6827, Standard deviation: 423.3, CV(%): 6.2,
• Sample 1: n=24, Mean(pg/ml): 1522, Standard deviation: 123.3, CV(%): 8.1
• Sample 2: n=24, Mean(pg/ml): 3594, Standard deviation: 230, CV(%): 6.4
• Sample 3: n=24, Mean(pg/ml): 7133, Standard deviation: 492.2, CV(%): 6.9

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

References for Lipocalin 2 ELISA Kit (ABIN411401)

Lei, Li, Zeng, Mu, Yang, Zhou, Wang, Zhang: "Value of urinary KIM-1 and NGAL combined with serum Cys C for predicting acute kidney injury secondary to decompensated cirrhosis." in: Scientific reports, Vol. 8, Issue 1, pp. 7962, (2018) (PubMed).

Yigit, Ulu, Gozel, Taskapan, Ilhan, Dogukan: "Neutrophil gelatinase-associated lipocalin reflects the severity of anemia without iron deficiency and secondary hyperparathyroidism in hemodialysis patients." in: Northern clinics of Istanbul, Vol. 4, Issue 1, pp. 36-42, (2017) (PubMed).

Benli, Ayyildiz, Cirrik, Noyan, Ayyildiz, Cirakoglu: "Early term effect of ureterorenoscopy (URS) on the Kidney: research measuring NGAL, KIM-1, FABP and CYS C levels in urine." in: International braz j urol : official journal of the Brazilian Society of Urology, Vol. 43, Issue 5, pp. 887-895, (2017) (PubMed).

Farahbakhsh-Farsi, Djazayery, Eshraghian, Koohdani, Zarei, Javanbakht, Derakhshanian, Djalali: "Effect of Omega-3 Supplementation on Lipocalin 2 and Retinol-Binding Protein 4 in Type 2 Diabetic Patients." in: Iranian journal of public health, Vol. 45, Issue 2, pp. 179-85, (2016) (PubMed).

Libório, Braz, Seguro, Meneses, Neves, Pedrosa, Cavalcanti, Martins, Daher: "Endothelial glycocalyx damage is associated with leptospirosis acute kidney injury." in: The American journal of tropical medicine and hygiene, Vol. 92, Issue 3, pp. 611-6, (2015) (PubMed).

Liu, Dong, Sun, Liu: "A novel fluid resuscitation protocol: provide more protection on acute kidney injury during septic shock in rats." in: International journal of clinical and experimental medicine, Vol. 7, Issue 4, pp. 919-26, (2014) (PubMed).

Merrikhi, Gheissari, Mousazadeh: "Urine and serum neutrophil gelatinase-associated lipocalin cut-off point for the prediction of acute kidney injury." in: Advanced biomedical research, Vol. 3, pp. 66, (2014) (PubMed).

Chen, Lu, Wang, Bian, Cheng, Su, Zhou: "Elevated urinary levels of cystatin C and neutrophil gelatinase-associated lipocalin in Henoch-Schönlein purpura patients with renal involvement." in: PLoS ONE, Vol. 9, Issue 6, pp. e101026, (2014) (PubMed).

Yazdani, Merrikhi, Beni, Baradaran, Soleimani, Musazade: "Association between neutrophil geletinase-associated lipocalin and iron deficiency anemia in children on chronic dialysis." in: Journal of research in medical sciences : the official journal of Isfahan University of Medical Sciences, Vol. 19, Issue 7, pp. 624-8, (2014) (PubMed).

Koca, Olguner, Ergür, Altekin, Ta?dö?en, Duru, Girgin, Gündüz, Cilaker M?c?l?, Güzelda?, Akku?: "The effects of dexmedetomidine on secondary acute lung and kidney injuries in the rat model of intra-abdominal sepsis." in: TheScientificWorldJournal, Vol. 2013, pp. 292687, (2013) (PubMed).

Sun, Meng, Jiang, Liu, Lei, Su, Duan, Wu, Xia, Xia: "Protective effect of ginsenoside Rb1 against intestinal ischemia-reperfusion induced acute renal injury in mice." in: PLoS ONE, Vol. 8, Issue 12, pp. e80859, (2013) (PubMed).

Olguner, Koca, Altekin, Ergür, Duru, Girgin, Ta?dö?en, Gündüz, Güzelda?, Akku?, Micili: "Ischemic preconditioning attenuates lipid peroxidation and apoptosis in the cecal ligation and puncture model of sepsis." in: Experimental and therapeutic medicine, Vol. 5, Issue 6, pp. 1581-1588, (2013) (PubMed).

Target Details for Lipocalin 2

Target: Lipocalin 2 (LCN2)

Alternative Name: LCN2 (LCN2 Products)

Synonyms: LCN2 ELISA Kit, 24p3 ELISA Kit, MSFI ELISA Kit, NGAL ELISA Kit, AW212229 ELISA Kit, Sip24 ELISA Kit, lipocalin 2 ELISA Kit, LCN2 ELISA Kit, Lcn2 ELISA Kit

Background:

Protein Function: Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5- DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation: iron-loaded form increases intracellular iron concentration without promoting apoptosis, while iron-free form decreases intracellular iron levels, inducing expression of the proapoptotic protein BCL2L11/BIM, resulting in apoptosis. Involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth. .

Background: Lipocalin-2(LCN2), also known as NGAL, is a protein associated with neutrophil gelatinase.1 The LCN2 gene is located at 9q34 and contains 7 exons.2 The 25-kD LCN2 protein is believed to bind small lipophilic substances such as bacteria-derived lipopolysaccharide(LPS) and formylpeptides and may function as a modulator of inflammation. NGAL tightly binds bacterial catecholate-type ferric siderophores through a cyclically permuted, hybrid electrostatic/cation-pi interaction and is a potent bacteriostatic agent in iron-limiting conditions.3 The primary LCN2 transcript is 3,696 nucleotides long, and the processed transcript is 809 nucleotides long.4 LCN2 expression in adult bone marrow, uterus, prostate, salivary gland, stomach, appendix, colon, trachea, and lung, and in fetal spleen and lung.The standard product used in this kit is recombinant human NGAL, consisting of 178 amino acids with the molecular mass of 22KDa.

Synonyms: Neutrophil gelatinase-associated lipocalin,NGAL,25 kDa alpha-2-microglobulin-related subunit of MMP-9,Lipocalin-2,Oncogene 24p3,Siderocalin LCN2,p25,LCN2,HNL, NGAL,

Full Gene Name: Neutrophil gelatinase-associated lipocalin

Cellular Localisation: Secreted . Upon binding to the SLC22A17 (24p3R) receptor, it is internalized.

Gene ID: 3934

UniProt: P80188

Pathways: Cellular Response to Molecule of Bacterial Origin, Transition Metal Ion Homeostasis