One Step Polymer HRP Mouse and Rat IgG (H+L)
IBSC One-Step anti-mouse and rat IgG (H+L); biotin/avidin free system stains membranes, cytoplasmic and nuclear antigens. It provides the user with a rapid and easy to use IHC detection system.
One Step Polymer HRP Mouse and Rat IgG (H+L)
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Ships: 1-2 business days
IHC/ICC procedure for frozen, paraffin sections and cell smears:
- Deparafinize and hydrate tissue sections through xylene or other clearing agents and graded alcohols.(For frozen sections or cell smears; use unfixed, acetone fixed or appropriate fixative for the antigen in question; for cell smears it may be necessary to permealize the cell by detergent, please refer to antibody protocol).
- Wash 2-3 with distilled or deionized water.
- Incubate sections/cell smear with Endoblocker (#1) for 5-10 minutes at room temperature (RT). Wash with distilled water 3X. Note: If antigen retriever (Trypsin AR-6541, Pronase AR-6542, Pepsin AR-6543, Citrate buffer AR-6544, Buffer w EDTA pH 8.5 AR-6545, Tris buffer pH 10 AR-6546) is required it can be applied at this step. Please refer to data sheet for the primary antibody.
- Wash slide with PBS or Tris saline buffer (with 0.02-0.05% nonionic detergent, Triton X100, Tween 20 or NP-40) or washing buffer (Immuno Automation buffer IBSC cat # AR-6561) 3X.
- Incubate sections/ cell smear in Protein blocking solution (#2), for 5-10 minutes at RT.
- Incubate sections/cell smear with primary antibody (NOT SUPPLIED, ONLY BUFFER IS SUPPLIED FOR DILUTION) for 20-30 minutes at RT. (For more information, refer to instructions for primary antibody). The primary antibody dilution buffer supplied can also be used as a negative control.
- Wash slide with PBS 5-7X.
- Incubate with One-Step HRP polymer (#4) for 20-30 minutes at RT.
- Wash slide 5-7 times with buffer. Caution: Peroxidase reagents are destroyed by sodium azide and should be avoided in all buffers and regents.
- Wash slide with deionized or distilled for 2-3X.
- Incubate with AEC or DAB chromogen reagent (Not supplied).
- Wash 5-7X with buffer.
- Incubate with counterstain (Not supplied).
- Wash slide with tap water distilled water.
- Mount slide with appropriate mounting medium.
Reagent Provided: 15 ml = 150 tests and 50 ml = 500 tests when 0.1 ml is applied per slide
IH-8061-15:Washing buffer, antigen retrievers, positive or negative control, primary antibody, chromogen, counterstain and mounting medium
IH-8061-15:
- Ready-to-use, Peroxidase Block, Hydrogen Peroxide, 15 ml (White cap)
- Ready-to-use Protein Blocking Solution, 15 ml (Blue cap)
- Primary Antibody Dilution Buffer green color, 30 ml (for dilution of primary antibody). Please refer to the data sheet of primary antibody for dilution. Use this buffer as a Negative Control
- One-step HRP-anti-Mouse and Rat Polymer, 15 ml (Orange cap)
- Ready-to-use, Peroxidase Block, Hydrogen Peroxide, 50 ml (White cap)
- Ready-to-use Protein Blocking Solution, 50 ml (Blue cap)
- Primary Antibody Dilution Buffer green color, 30 ml (for dilution of primary antibody). Please refer to the data sheet of primary antibody for dilution. Use this buffer as a Negative Control
- One-step HRP-anti-Mouse and Rat Polymer, 50 ml (Orange cap)