The Rat neutrophil gelatinase-associated lipocalin (NGAL) ELISA Kit is used to quantitatively measure NGAL concentrations in rat serum, plasma, urine, or tissue homogenates. It performs well in important characteristics, including sensitivity, specificity, precision, recovery, linearity, and lot-to-lot consistency. This assay is based on the sandwich ELISA mechanism and enzyme-substrate chromogenic reaction. The solution color develops proportionally to the amount of NGAL in the sample. And the intensity of the color can be measured at 450 nm via a microplate reader.
NGAL was originally isolated from secondary granules of human neutrophils as a 25 KDa protein covalently linked to matrix metalloproteinase-9 (MMP-9) in human neutrophils. It is highly induced in the kidney after ischemic or nephrotoxic acute kidney injury (AKI) in animal models. NGAL has been reported as an early urine and plasma biomarker of AKI in pediatric and adult cardiac surgery. Urinary NGAL has been employed as a routine measurement in AKI patients by King Chulalongkorn Memorial Hospital since 2015.
||Lcn2 ELISA Kit; Neutrophil gelatinase-associated lipocalin ELISA Kit; NGAL ELISA Kit; Alpha-2-microglobulin-related protein ELISA Kit; Alpha-2U globulin-related protein ELISA Kit; Lipocalin 24p3 ELISA Kit; Lipocalin-2 ELISA Kit; Siderocalin LCN2 ELISA Kit; p25 ELISA Kit
||Rattus norvegicus (Rat)
||serum, plasma, urine, tissue homogenates
||0.312 ng/mL-20 ng/mL
|Intra-assay Precision (Precision within an assay): CV%<8%|| || || |
|Three samples of known concentration were tested twenty times on one plate to assess. || |
|Inter-assay Precision (Precision between assays): CV%<10%|| || || |
|Three samples of known concentration were tested in twenty assays to assess. || || |
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|To assess the linearity of the assay, samples were spiked with high concentrations of rat NGAL in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.|
| ||Sample||Serum(n=4)|| |
|1:1||Average %||88|| |
|Range %||80-98|| |
|1:2||Average %||95|| |
|Range %||89-99|| |
|1:4||Average %||85|| |
|Range %||81-90|| |
|1:8||Average %||104|| |
|Range %||99-107|| |
|The recovery of rat NGAL spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.|
|Sample Type||Average % Recovery||Range|| |
|Serum (n=5) ||98||93-104|| |
|EDTA plasma (n=4)||95||90-100|| |
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|These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|20||2.451 ||2.498 ||2.475 ||2.287 || |
|10||1.954 ||1.967 ||1.961 ||1.773 || |
|5||1.226 ||1.269 ||1.248 ||1.060 || |
|2.5||0.758 ||0.789 ||0.774 ||0.586 || |
|1.25||0.529 ||0.505 ||0.517 ||0.329 || |
|0.625||0.380 ||0.351 ||0.366 ||0.178 || |
|0.312||0.279 ||0.271 ||0.275 ||0.087 || |
|0||0.187 ||0.189 ||0.188 || || |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-rat NGAL. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled NGAL antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent(50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
|Materials not provided
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Test tubes for dilution
|ELISA kit FAQs
||Store at 2-8°C. Please refer to protocol.
||3-5 working days