MUC5AC ELISA Kit

Catalog No. ABIN6574198

Product Details MUC5AC ELISA Kit

Target: MUC5AC (Mucin 5AC, Oligomeric Mucus/gel-Forming (MUC5AC))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 78 pg/mL - 5000 pg/mL

Minimum Detection Limit: 78 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of MUC5AC in human tissue homogenates, cell lysates, cell culture supernates.

We offer validation data (WB) for 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: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Mucin 5 Subtype AC (MUC5AC)

Cross-Reactivity (Details): No significant cross-reactivity or interference between Mucin 5 Subtype AC (MUC5AC) and analogues was observed.

Sensitivity: 30 pg/mL

Components:

• 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
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

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

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. 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.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 10,000pg/mL. Firstly dilute the stock solution to 5,000pg/mL and the diluted standard serves as the highest standard (5,000pg/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 5,000pg/mL, 2,500pg/mL, 1,250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL, 78pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). 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.
4. 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).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. 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.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

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%

Restrictions: For Research Use only

Handling

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.

Storage: 4 °C/-20 °C

Storage Comment:

1. 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.
2. 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.

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Expiry Date: 6 months

References for MUC5AC ELISA Kit (ABIN6574198)

Shah, Ishinaga, Takeuchi: "Clarithromycin inhibits TNF-α-induced MUC5AC mucin gene expression via the MKP-1-p38MAPK-dependent pathway." in: International immunopharmacology, Vol. 49, pp. 60-66, (2018) (PubMed).

Reuter, Alzheimer, Walles, Oelschlaeger: "An adherent mucus layer attenuates the genotoxic effect of colibactin." in: Cellular microbiology, Vol. 20, Issue 2, (2018) (PubMed).

Miyake, Mori, Mano, Imanaka, Nakamura: "Development of a highly sensitive and reliable enzyme-linked immunosorbent assay for MUC5AC in human tears extracted from Schirmer strips." in: Clinical ophthalmology (Auckland, N.Z.), Vol. 12, pp. 1571-1580, (2018) (PubMed).

Ma, Ma: "Plantamajoside Inhibits Lipopolysaccharide-Induced MUC5AC Expression and Inflammation through Suppressing the PI3K/Akt and NF-κB Signaling Pathways in Human Airway Epithelial Cells." in: Inflammation, Vol. 41, Issue 3, pp. 795-802, (2018) (PubMed).

Guan, Zhu, Shen, Jia, Jin, Dong, Xie: "Salvianolic acid B improves airway hyperresponsiveness by inhibiting MUC5AC overproduction associated with Erk1/2/P38 signaling." in: European journal of pharmacology, Vol. 824, pp. 30-39, (2018) (PubMed).

Shah, Ishinaga, Takeuchi: "Oxytetracycline Inhibits Mucus Secretion and Inflammation in Human Airway Epithelial Cells." in: Chemotherapy, Vol. 62, Issue 5, pp. 301-306, (2017) (PubMed).

Jiang, Wang, Tang, Yao, Zhou: "Regulation of Viral Infection-induced Airway Remodeling Cytokine Production by the TLR3-EGFR Signaling Pathway in Human Bronchial Epithelial Cells." in: COPD, pp. 1-6, (2016) (PubMed).

Uchino, Uchino, Yokoi, Dogru, Kawashima, Komuro, Sonomura, Kato, Argüeso, Kinoshita, Tsubota: "Impact of Cigarette Smoking on Tear Function and Correlation between Conjunctival Goblet Cells and Tear MUC5AC Concentration in Office Workers." in: Scientific reports, Vol. 6, pp. 27699, (2016) (PubMed).

Ishinaga, Kitano, Toda, DAlessandro-Gabazza, Gabazza, Shah, Takeuchi: "Interleukin-33 induces mucin gene expression and goblet cell hyperplasia in human nasal epithelial cells." in: Cytokine, Vol. 90, pp. 60-65, (2016) (PubMed).

Shirasaki, Kanaizumi, Seki, Himi: "Leukotriene E4 induces MUC5AC release from human airway epithelial NCI-H292 cells." in: Allergology international : official journal of the Japanese Society of Allergology, Vol. 64, Issue 2, pp. 169-74, (2015) (PubMed).

Sibila, Suarez-Cuartin, Rodrigo-Troyano, Fardon, Finch, Mateus, Garcia-Bellmunt, Castillo, Vidal, Sanchez-Reus, Restrepo, Chalmers: "Secreted mucins and airway bacterial colonization in non-CF bronchiectasis." in: Respirology (Carlton, Vic.), Vol. 20, Issue 7, pp. 1082-8, (2015) (PubMed).

Ruzzenente, Iacono, Conci, Bertuzzo, Salvagno, Ruzzenente, Campagnaro, Valdegamberi, Pachera, Bagante, Guglielmi: "A novel serum marker for biliary tract cancer: diagnostic and prognostic values of quantitative evaluation of serum mucin 5AC (MUC5AC)." in: Surgery, Vol. 155, Issue 4, pp. 633-9, (2014) (PubMed).

Danese, Ruzzenente, Ruzzenente, Iacono, Bertuzzo, Gelati, Conci, Bendinelli, Bonizzato, Guglielmi, Salvagno, Lippi, Guidi: "Assessment of bile and serum mucin5AC in cholangiocarcinoma: diagnostic performance and biologic significance." in: Surgery, Vol. 156, Issue 5, pp. 1218-24, (2014) (PubMed).

Musrap, Karagiannis, Saraon, Batruch, Smith, Diamandis: "Proteomic analysis of cancer and mesothelial cells reveals an increase in Mucin 5AC during ovarian cancer and peritoneal interaction." in: Journal of proteomics, Vol. 103, pp. 204-15, (2014) (PubMed).

Zhang, Zhu, Yang, Pan, Jiang, Sun, Wang, Xiao: "The human Cathelicidin LL-37 induces MUC5AC mucin production by airway epithelial cells via TACE-TGF-?-EGFR pathway." in: Experimental lung research, Vol. 40, Issue 7, pp. 333-42, (2014) (PubMed).

Hao, Kuang, Xu, Walling, Lau: "Pyocyanin-induced mucin production is associated with redox modification of FOXA2." in: Respiratory research, Vol. 14, Issue 1, pp. 82, (2013) (PubMed).

Zhang, Jiang, Sun, Wang, Yang, Pan, Zhu, Xiao: "The human cathelicidin LL-37 enhances airway mucus production in chronic obstructive pulmonary disease." in: Biochemical and biophysical research communications, Vol. 443, Issue 1, pp. 103-9, (2013) (PubMed).

Nie, Wu, Li, Xie, Luo, Shen, Su: "Naringin attenuates EGF-induced MUC5AC secretion in A549 cells by suppressing the cooperative activities of MAPKs-AP-1 and IKKs-I?B-NF-?B signaling pathways." in: European journal of pharmacology, Vol. 690, Issue 1-3, pp. 207-13, (2012) (PubMed).

Maker, Katabi, Gonen, DeMatteo, DAngelica, Fong, Jarnagin, Brennan, Allen: "Pancreatic cyst fluid and serum mucin levels predict dysplasia in intraductal papillary mucinous neoplasms of the pancreas." in: Annals of surgical oncology, Vol. 18, Issue 1, pp. 199-206, (2011) (PubMed).

Katti, Sathyanesan: "Changes in the hypothalamo-neurohypophysial complex of lead treated teleostean fish Clarias batrachus (L.)." in: Zeitschrift für mikroskopisch-anatomische Forschung, Vol. 100, Issue 3, pp. 347-52, (1986) (PubMed).

Target Details for MUC5AC

Target: MUC5AC (Mucin 5AC, Oligomeric Mucus/gel-Forming (MUC5AC))

Alternative Name: Mucin 5 Subtype AC (MUC5AC) (MUC5AC Products)

Synonyms: MUC5 ELISA Kit, TBM ELISA Kit, leB ELISA Kit, 2210005L13Rik ELISA Kit, MGM ELISA Kit, mucin 5AC, oligomeric mucus/gel-forming ELISA Kit, putative gastric mucin ELISA Kit, mucin 5, subtypes A and C, tracheobronchial/gastric ELISA Kit, mucin 2, oligomeric mucus/gel-forming ELISA Kit, MUC5AC ELISA Kit, Smp_144070 ELISA Kit, Muc5ac ELISA Kit, MUC2 ELISA Kit

UniProt: P98088