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Human FABP4 ELISA Kit

SKU:
HUFI00539
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P15090
Sensitivity:
0.234ng/ml
Range:
0.391-25ng/ml
ELISA Type:
Sandwich
Synonyms:
FABP4, A-FABP, AFABP, ALBP, aP2, Adipocyte lipid-binding protein, Adipocyte-type fatty acid-binding protein, Fatty acid-binding protein 4, fatty acid-binding protein, adipocyte
Reactivity:
Human
Research Area:
Signal Transduction
€499
Frequently bought together:

Description

Human FABP4 ELISA Kit

The Human FABP4 ELISA Kit is specifically designed for the precise detection of Fatty Acid Binding Protein 4 levels in human serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures accurate and consistent results, making it an excellent tool for various research purposes.Fatty Acid Binding Protein 4, also known as adipocyte protein 2 (aP2), is a key regulator of lipid metabolism and inflammation.

It plays a critical role in conditions such as obesity, diabetes, and cardiovascular diseases, making it a valuable biomarker for studying these diseases and potential therapeutic interventions. Overall, the Human FABP4 ELISA Kit is a valuable asset for researchers looking to study the role of FABP4 in various physiological and pathological processes, with the potential for identifying novel therapeutic targets and strategies.

Product Name:Human FABP4 ELISA Kit
Product Code:HUFI00539
Size:96 Assays
Alias:FABP4, A-FABP, AFABP, ALBP, aP2, Adipocyte lipid-binding protein, Adipocyte-type fatty acid-binding protein, Fatty acid-binding protein 4, fatty acid-binding protein, adipocyte
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human FABP4 concentrations in serum plasma and other biological fluids.
Sensitivity:0.234ng/ml
Range:0.391-25ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human FABP4 and the recovery rates were calculated by comparing the measured value to the expected amount of Human FABP4 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10496
EDTA plasma(n=5)85-9992
UFH plasma(n=5)92-10397
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human FABP4 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-97%88-105%92-105%
EDTA plasma(n=5)88-91%84-95%82-97%
UFH plasma(n=5)85-99%83-100%84-97%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP15090
UniProt Protein Function:FABP4: a lipid transport protein that is the predominant fatty acid binding protein found in adipocytes. Also expressed in macrophages. Binds both long chain fatty acids and retinoic acid. Delivers long-chain fatty acids and retinoic acid to their cognate receptors in the nucleus. Homodimer. Forms a beta-barrel structure that accommodates hydrophobic ligands in its interior. Interacts with PPARG. Monomer. FABP4 knockout mice fed a high-fat and high-calorie diet become obese but develop neither insulin resistance nor diabetes, suggesting that this protein might be a link between obesity and insulin resistance and diabetes. Mice deficient in both FABP4 and ApoE show protection against atherosclerosis when compared with mice deficient only in ApoE. Mice carrying a FABP4 genetic variant exhibit both reduced FABP4 expression and a reduced potential for developing type 2 diabetes and coronary heart disease. A related study in humans indicated a similar pattern, suggesting that FABP4 may be a potential therapeutic target in the treatment of these disorders. Belongs to the calycin superfamily,fatty-acid binding protein (FABP) family.
UniProt Protein Details:Protein type:Lipid-binding

Chromosomal Location of Human Ortholog: 8q21

Cellular Component: cytoplasm; lipid particle; nucleus

Molecular Function:transporter activity; fatty acid binding

Biological Process: cholesterol homeostasis; transport; white fat cell differentiation; triacylglycerol catabolic process; brown fat cell differentiation; negative regulation of protein kinase activity; cytokine production; negative regulation of transcription, DNA-dependent; positive regulation of inflammatory response

NCBI Summary:FABP4 encodes the fatty acid binding protein found in adipocytes. Fatty acid binding proteins are a family of small, highly conserved, cytoplasmic proteins that bind long-chain fatty acids and other hydrophobic ligands. It is thought that FABPs roles include fatty acid uptake, transport, and metabolism. [provided by RefSeq, Jul 2008]
UniProt Code:P15090
NCBI GenInfo Identifier:119781
NCBI Gene ID:2167
NCBI Accession:P15090.3
UniProt Secondary Accession:P15090,Q6IBA1,
UniProt Related Accession:P15090
Molecular Weight:132
NCBI Full Name:Fatty acid-binding protein, adipocyte
NCBI Synonym Full Names:fatty acid binding protein 4, adipocyte
NCBI Official Symbol:FABP4  
NCBI Official Synonym Symbols:aP2; ALBP; AFABP; A-FABP; HEL-S-104  
NCBI Protein Information:fatty acid-binding protein, adipocyte; fatty acid-binding protein 4; adipocyte lipid-binding protein; epididymis secretory protein Li 104; adipocyte-type fatty acid-binding protein
UniProt Protein Name:Fatty acid-binding protein, adipocyte
UniProt Synonym Protein Names:Adipocyte lipid-binding protein; ALBP; Adipocyte-type fatty acid-binding protein; A-FABP; AFABP; Fatty acid-binding protein 4
Protein Family:Fatty acid-binding protein
UniProt Gene Name:FABP4  
UniProt Entry Name:FABP4_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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