HMOX1 ELISA Kit

Catalog No. ABIN6963168

Product Details HMOX1 ELISA Kit

Target: HMOX1 (Heme Oxygenase (Decycling) 1 (HMOX1))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.16 ng/mL - 10 ng/mL

Minimum Detection Limit: 0.16 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

Specificity: This kit recognizes Rat HO1 in samples. No Significant cross-reactivity or interference between Rat HO1 and analogues was observed.

Sensitivity: 0.1 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 100 μL

Assay Time: 3.5 h

Plate: Pre-coated

Protocol:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Reagent Preparation:

1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 10 ng/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 10, 5, 2.5, 1.25, 0.63, 0.32, 0.16, 0 ng/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 10 ng/mL working solution to the first tube and mix up to produce a 5 ng/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.

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, mid range and high level Rat HO1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat HO1 were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 6 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

Expiry Date: 6 months

References for HMOX1 ELISA Kit (ABIN6963168)

Balaha, Alamer, Eisa, Aljohani: "Shikonin Alleviates Gentamicin-Induced Renal Injury in Rats by Targeting Renal Endocytosis, SIRT1/Nrf2/HO-1, TLR-4/NF-κB/MAPK, and PI3K/Akt Cascades." in: Antibiotics (Basel, Switzerland), Vol. 12, Issue 5, (2023) (PubMed).

Balaha, Almalki, Alahmari, Ahmed, Balaha: "AMPK/mTOR-driven autophagy & Nrf2/HO-1 cascade modulation by amentoflavone ameliorates indomethacin-induced gastric ulcer." in: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, Vol. 151, pp. 113200, (2022) (PubMed).

Gao, Hou, Wang, Guo, Yuan, Jiang, Duan, Zhang, Xu, Che, Sun, Li, Zhang, Sun, Li: "Octreotide alleviates pancreatic damage caused by paraquat in rats by reducing inflammatory responses and oxidative stress." in: Environmental toxicology and pharmacology, Vol. 80, pp. 103456, (2021) (PubMed).

Mahran: "New insights into the protection of growth hormone in cisplatin-induced nephrotoxicity: The impact of IGF-1 on the Keap1-Nrf2/HO-1 signaling." in: Life sciences, Vol. 253, pp. 117581, (2020) (PubMed).

Abdelaziz, Mantawy, Gad, Fawzy, Azab: "Activation of pCREB/Nrf-2 signaling mediates re-positioning of liraglutide as hepato-protective for methotrexate -induced liver injury (MILI)." in: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association, Vol. 132, pp. 110719, (2019) (PubMed).

Target Details for HMOX1

Target: HMOX1 (Heme Oxygenase (Decycling) 1 (HMOX1))

Alternative Name: Heme Oxygenase 1 (HMOX1 Products)

Synonyms: HMOX1 ELISA Kit, ARABIDOPSIS THALIANA HEME OXYGENASE 1 ELISA Kit, ATHO1 ELISA Kit, F18A8.4 ELISA Kit, F18A8_4 ELISA Kit, GENOMES UNCOUPLED 2 ELISA Kit, GUN2 ELISA Kit, HEME OXYGENASE ELISA Kit, HEME OXYGENASE 1 ELISA Kit, HEME OXYGENASE 6 ELISA Kit, HO1 ELISA Kit, HY1 ELISA Kit, HY6 ELISA Kit, PLASTID HEME OXYGENASE ELISA Kit, REVERSAL OF THE DET PHENOTYPE 4 ELISA Kit, HO-1 ELISA Kit, wu:fc27c04 ELISA Kit, zgc:65984 ELISA Kit, D8Wsu38e ELISA Kit, Hemox ELISA Kit, Hmox ELISA Kit, Hsp32 ELISA Kit, HEOXG ELISA Kit, Heox ELISA Kit, Ho-1 ELISA Kit, Ho1 ELISA Kit, hsp32 ELISA Kit, MGC132176 ELISA Kit, Hmox1 ELISA Kit, HMOX1D ELISA Kit, HSP32 ELISA Kit, bK286B10 ELISA Kit, heme oxygenase 1 ELISA Kit, Plant heme oxygenase (decyclizing) family protein ELISA Kit, Heme oxygenase ELISA Kit, heme oxygenase ELISA Kit, heme oxygenase 1, chloroplastic ELISA Kit, heme oxygenase 1a ELISA Kit, heme oxygenase 1 L homeolog ELISA Kit, HMOX1 ELISA Kit, TED4 ELISA Kit, ho1 ELISA Kit, P9301_RS17555 ELISA Kit, A9601_RS17590 ELISA Kit, MAE_RS06565 ELISA Kit, HO1 ELISA Kit, hmox1a ELISA Kit, CPE0214 ELISA Kit, Cyan7425_2962 ELISA Kit, Hmox1 ELISA Kit, hmox1.L ELISA Kit, AM1_C0205 ELISA Kit, CC1G_11686 ELISA Kit, CpipJ_CPIJ007246 ELISA Kit

Background: HO1, HMOX1, HMOX1D, HSP32, Bk286b10

Pathways: Transition Metal Ion Homeostasis, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, SARS-CoV-2 Protein Interactome