WNT5A ELISA Kit (Wingless-Type MMTV Integration Site Family, Member 5A) ELISA Kit
- Human, Mouse, Rat
- Detection Method
- Method Type
- Sandwich ELISA
- Detection Range
- 0.15 ng/mL - 10 ng/mL
- Minimum Detection Limit
- 0.15 ng/mL
The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of WNT5A in tissue homogenates, cell lysates, cell culture supernates. Due to the 98% homology of the sequence among different species, the kit can be used to detect human, mouse and rat samples.
We offer validation data (WB) for each of the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.
- Sample Type
- Cell Culture Supernatant, Cell Lysate, Tissue Homogenate
- Analytical Method
- This assay has high sensitivity and excellent specificity for detection of Wingless Type MMTV Integration Site Family, Member 5A (WNT5A)
- Cross-Reactivity (Details)
- No significant cross-reactivity or interference between Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) and analogues was observed.
- 0.059 ng/mL
- Pre-coated, ready to use 96-well strip plate, flat buttom
- Plate sealer for 96 wells
- Reference Standard
- Standard Diluent
- Detection Reagent A
- Detection Reagent B
- Assay Diluent A
- Assay Diluent B
- TMB Substrate
- Stop Solution
- Wash Buffer (30 x concentrate)
- Instruction manual
- Material not included
- Microplate reader with 450 ± 10nm filter.
- Precision single or multi-channel pipettes and disposable tips.
- Microcentrifuge tubes for diluting samples.
- Deionized or distilled water.
- Absorbent paper for blotting the microtiter plate.
- Container for Wash Solution
- Incubator capable of maintaining 37 °C.
- 0.01mol/L (or 1×) Phosphate Buffered Saline (PBS), pH7.0-7.2.
Wingless-Type MMTV Integration Site Family, Member 5A (WNT5A) ELISA Kit ELISA Kit
WNT5A Reactivity: Mouse Colorimetric Sandwich ELISA 0.15 ng/mL - 10 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.
Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.
Information on antibodies:
The provided antibodies and their host vary in different kits.
- Sample Volume
- 100 μL
- Assay Time
- 3 h
- Prepare all reagents, samples and standards,
- Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
- Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
- Aspirate and wash 3 times,
- Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
- Aspirate and wash 5 times,
- Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
- Add 50μL Stop Solution. Read at 450nm immediately.
- Reagent Preparation
- Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
- Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 80 ng/mL. Please firstly dilute the stock solution to 10 ng/mL and the diluted standard serves as the highest standard (10 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL.
- Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
- Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
- TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
- Making serial dilution in the wells directly is not permitted.
- Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
- Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
- The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
- If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
- Contaminated water or container for reagent preparation will influence the detection result.
- Assay Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%
- For Research Use only
- Precaution of Use
- The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
- 4 °C/-20 °C
- Storage Comment
- For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
- For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
It is strongly recommended to use the remaining reagents within 1 month, if this is done before the expiry date of the kit. Please refer to the label on the kit packaging for the expiration date of the kit. All components are stable until the expiration date.
- Expiry Date
- 6 months
Cho, Kang, Lee, Lee, Seol, Lee, Park, Jung: "Effect of SFRP5 (Secreted Frizzled-Related Protein 5) on the WNT5A (Wingless-Type Family Member 5A)-Induced Endothelial Dysfunction and Its Relevance With Arterial Stiffness in Human Subjects." in: Arteriosclerosis, thrombosis, and vascular biology, Vol. 38, Issue 6, pp. 1358-1367, 2018 (PubMed).
Schulte, Kragelund, Müller, Hagen, Elke, Titz, Schädler, Schumacher, Weiler, Bewig, Schreiber, Laudes: "The wingless-related integration site-5a/secreted frizzled-related protein-5 system is dysregulated in human sepsis." in: Clinical and experimental immunology, Vol. 180, Issue 1, pp. 90-7, 2015 (PubMed).
Chatani, Kamada, Kizu, Ogura, Furuta, Egawa, Hamano, Ezaki, Kiso, Shimono, Ouchi, Yoshida, Takehara: "Secreted frizzled-related protein 5 (Sfrp5) decreases hepatic stellate cell activation and liver fibrosis." in: Liver international : official journal of the International Association for the Study of the Liver, Vol. 35, Issue 8, pp. 2017-26, 2015 (PubMed).
Xu, Xu, Zhao, Yang, Liu, Wen, Zhang: "Wnt5a reverses the inhibitory effect of hyperoxia on transdifferentiation of alveolar epithelial type II cells to type I cells." in: Journal of physiology and biochemistry, Vol. 71, Issue 4, pp. 823-38, 2015 (PubMed).
Bougault, Briolay, Boutet, Pilet, Delplace, Le Goff, Guicheux, Blanchard, Magne: "Wnt5a is expressed in spondyloarthritis and exerts opposite effects on enthesis and bone in murine organ and cell cultures." in: Translational research : the journal of laboratory and clinical medicine, 2015 (PubMed).
Prats-Puig, Soriano-Rodríguez, Carreras-Badosa, Riera-Pérez, Ros-Miquel, Gomila-Borja, de Zegher, Ibáñez, Bassols, López-Bermejo: "Balanced duo of anti-inflammatory SFRP5 and proinflammatory WNT5A in children." in: Pediatric research, Vol. 75, Issue 6, pp. 793-7, 2014 (PubMed).
Tan, Wang, Chu, Liu, Yi, Xiao: "SFRP5 correlates with obesity and metabolic syndrome and increases after weight loss in children." in: Clinical endocrinology, Vol. 81, Issue 3, pp. 363-9, 2014 (PubMed).
Catalán, Gómez-Ambrosi, Rodríguez, Pérez-Hernández, Gurbindo, Ramírez, Méndez-Giménez, Rotellar, Valentí, Moncada, Martí, Sola, Silva, Salvador, Frühbeck: "Activation of noncanonical Wnt signaling through WNT5A in visceral adipose tissue of obese subjects is related to inflammation." in: The Journal of clinical endocrinology and metabolism, Vol. 99, Issue 8, pp. E1407-17, 2014 (PubMed).
Carstensen, Herder, Kempf, Erlund, Martin, Koenig, Sundvall, Bidel, Kuha, Roden, Tuomilehto: "Sfrp5 correlates with insulin resistance and oxidative stress." in: European journal of clinical investigation, Vol. 43, Issue 4, pp. 350-7, 2013 (PubMed).
Schulte, Müller, Neumann, Oberhäuser, Faust, Güdelhöfer, Brandt, Krone, Laudes: "Pro-inflammatory wnt5a and anti-inflammatory sFRP5 are differentially regulated by nutritional factors in obese human subjects." in: PLoS ONE, Vol. 7, Issue 2, pp. e32437, 2012 (PubMed).
- Cho, Kang, Lee, Lee, Seol, Lee, Park, Jung: "Effect of SFRP5 (Secreted Frizzled-Related Protein 5) on the WNT5A (Wingless-Type Family Member 5A)-Induced Endothelial Dysfunction and Its Relevance With Arterial Stiffness in Human Subjects." in: Arteriosclerosis, thrombosis, and vascular biology, Vol. 38, Issue 6, pp. 1358-1367, 2018 (PubMed).
- Alternative Name
- Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) (WNT5A ELISA Kit Abstract)
- hWNT5A, wnt-5A, wnt5a-B, WNT5A, 8030457G12Rik, Wnt-5a, Wnt family member 5A, Wnt family member 5A S homeolog, wingless-type MMTV integration site family, member 5a, wingless-type MMTV integration site family, member 5A, wingless-related MMTV integration site 5A, Wnt signaling ligand, WNT5A, Wnt5a, wnt5a.S, wnt5a, LOC578814, wnt5
- WNT Signaling, Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Regulation of Cell Size, Tube Formation