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2-step plus Poly-HRP Anti Mouse/Rabbit IgG Detection System (with DAB solution)

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      Catalog number:E-IR-R217

      Size:
      • 3mL
      • 6mL
      • 18mL
      Qty:
      - +
      Price: $60

      Lead Time:Order now, ship in 3 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Introduction

      2-step plus is a two-step immunohistochemical broad spectrum detection reagent. It polymerizes monovalent Fab fragments of secondary antibody and enzyme, which replaces the secondary antibody and tertiary antibody in traditional method, can directly amplify the binding signal of antibody-antigen. This method not only retains the specific binding ability of antibody with antigen, but also can effectively avoid space steric hindrance caused by excessive polymer molecules. Compared with the traditional SP three-step method, this kit has the characteristics of simple, rapid, high-sensitivity. This system abandons the using of biotin, so it can avoid background staining by endogenous biotin. It can be used in IHC, in which the primary antibody is monoclonal or polyclonal antibody derived from mouse or rabbit.

      Diluent for DAB  concentrated solution has been provided in this kit to avoid the influence of the different water acidity and alkalinity on the DAB Chromogenic Agent.


      Components

      Cat

      product

      3 mL

      6 mL

      18 mL

      Storage

      E-IR-R217A

      Normal Goat Blocking Buffer (Ready-to-Use)

      3 mL

      6 mL

      18 mL

      2~8°C

      E-IR-R217B

      Polyperoxidase-anti-Mouse/Rabbit IgG (Ready-to-Use)

      3 mL

      6 mL

      18 mL

      2~8°C

      E-IR-R217C

      3% H2O2

      3 mL

      6 mL

      18 mL

      2~8°C

      E-IR-R217D

      DAB Concentrate (20×)

      150 μL

      300 μL

      900 μL

      2~8°C

      E-IR-R217E

      DAB Substrate

      3 mL

      6 mL

      18 mL

      2~8°C

      Manual

      One Copy



      Sample dyeing

      1.  Dewax and hydrate the paraffin section.

      2.  Make thermal repair or digestion treatment to antigen of the tissue section if necessary according to antigen/antibody situation.

      3.  Incubate with E-IR-R217C (3% H2O2) for 10 min to eliminate endogenous peroxidase activity. Wash with PBS or TBS, 2 min×3 times.

      4.  Add E-IR-R217A (Normal Goat Blocking Buffer (Ready-to-Use)), incubate at 37°C for 30 min. Shake off any excess liquid.

      5.  Add primary antibody (From Mouse or Rabbit) with proper dilution ratio, incubate at 20~37°C for 1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS, 2 min×3 times. Dry the section with absorbent paper.

      6.  Add E-IR-R217B (Polyperoxidase-anti-Mouse/Rabbit IgG), incubate at room temperature or 37°C for 20 min. Wash with PBS or TBS, 2 min×3 times.

      7.  Add 1 drop (approximately 50 μL) of E-IR-R217D (DAB Concentrate) into each 1 mL of E-IR-R217E (DAB Substrate), mix fully and the mixed reagent is the DAB Working Solution. Prepare fresh solution before use and the prepared solution should be stored in the dark. Fresh prepared DAB Working Solution is valid within 4 hours and the unused solution must be abandoned

      8.  Take control of the DAB coloration period, the color of tan or brownish yellow is the positive signal. Avoid of excessive reaction.

      9.  Wash the section with deionized water terminate the chromogenic reaction, then operate the procedures of counterstaining, dehydrating, transparentizing and sealing

      Storage

      Store at 2~8°C, shading light. Avoid of freezing. Valid for 12 months. The reagents are valid within 6 months after opening.

      Citations

      1. T lymphocyte-derived extracellular vesicles aggravate abdominal aortic aneurysm by promoting macrophage lipid peroxidation and migration via pyruvate kinase muscle isozyme 2
        IF: 11.799
        Journal: Redox Biology(2022)
        DOI: 10.1016/j.redox.2022.102257
        PMID: 35149342
        Products: E-IR-R217 
        Sample type: aorta
        Reactivity: Mouse
      2. Syntenin Regulated by miR-216b Promotes Cancer Progression in Pancreatic Cancer
        IF: 6.244
        Journal: Frontiers in Oncology(2022)
        DOI: 10.3389/fonc.2022.790788
        Products: E-IR-R217 
      3. Naringin administration mitigates oxidative stress, anemia, and hypertension in lead acetate-induced cardio-renal dysfunction in cockerel chicks
        IF: 5.19
        Journal: ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH(2022)
        DOI: 10.1007/s11356-022-24656-4
        Products: E-IR-R217 
      4. L-arginine and lisinopril supplementation protects against sodium fluoride–induced nephrotoxicity and hypertension by suppressing mineralocorticoid receptor and angiotensin-converting enzyme 3 activity
        IF: 5.19
        Journal: ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH(2022)
        DOI: 10.1007/s11356-022-23784-1
        Products: E-IR-R217 
      5. Quantitative and qualitative analysis of pulmonary arterial hypertension fibrosis using wide-field second harmonic generation microscopy
        IF: 4.38
        Journal: Scientific Reports(2022)
        DOI: 10.1038/s41598-022-11473-5
        PMID: 35513702
        Products: E-IR-R217 
      6. Lymph Node Metastasis-Related Gene ITGA4 Promotes the Proliferation, Migration, and Invasion of Gastric Cancer Cells by Regulating Tumor Immune Microenvironment
        IF: 4.501
        Journal: Journal of Oncology(2022)
        DOI: 10.1155/2022/1315677
        PMID: 36254221
        Products: E-IR-R217 
        Sample type: GC tissues,paracancerous tissues
        Reactivity: Human
      7. Nigral and ventral tegmental area lesioning induces testicular and sperm morphological abnormalities in a rotenone model of Parkinson’s disease
        IF: 3.292
        Journal: ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY(2020)
        DOI: 10.1016/j.etap.2020.103412
        Products: E-IR-R217 
      8. FSIP1 Is Associated with Poor Prognosis and Can Be Used to Construct a Prognostic Model in Gastric Cancer
        IF: 3.434
        Journal: DISEASE MARKERS(2022)
        DOI: 10.1155/2022/2478551
        PMID: 35692888
        Products: E-IR-R217 
        Sample type: GC tissue
        Reactivity: Human
      9. Naringin abrogates angiotensin-converting enzyme (ACE) activity and podocin signaling pathway in cobalt chloride-induced nephrotoxicity and hypertension
        IF: 2.663
        Journal: BIOMARKERS(2022)
        DOI: 10.1080/1354750X.2022.2157489
        Products: E-IR-R217 
      10. Kolaviron ameliorates behavioural deficit and injury to striatal dopaminergic terminals via modulation of oxidative burden, DJ-1 depletion and CD45R + cells infiltration in MPTP-model of Parkinson’s disease
        IF: 2.726
        Journal: METABOLIC BRAIN DISEASE(2020)
        DOI: 10.1007/s11011-020-00578-3
        Products: E-IR-R217 
        Sample type: Tissue
        Reactivity: Mouse
      11. HBXIP protein overexpression predicts the poor prognosis of pancreatic ductal adenocarcinomas
        IF: 1.466
        Journal: PATHOLOGY RESEARCH AND PRACTICE(2018)
        DOI: 10.1016/j.prp.2018.12.016
        PMID: 30583814
        Products: E-IR-R217 
        Sample type: Tissue
        Reactivity: Human
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