Guinea Pig anti-Goat IgG Antibody
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- Target See all IgG products
- IgG
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Reactivity
- Goat
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Host
- Guinea Pig
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Clonality
- Polyclonal
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Application
- ELISA, Immunohistochemistry (IHC), Western Blotting (WB), Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Adaptor Antibody (AAb)
- Purpose
- Guinea pig anti goat IgG. Functions as an adapter to convert a primary antibody from one species into another.
- Specificity
- Specific for goat IgG
- Purification
- Purified
- Isotype
- IgG
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- Protocol
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Western Blot Protocol
- Run the SDS-PAGE at 250V.
- Soak the gel for 15 minutes in transfer buffer, then rinse with DI water.
- Soak the membrane for 5 minutes in DI water.
- Semi-dry transfer at 10V.
- After transfer, rinse the membrane once with PBS, then wash 3× for 5 minutes each with PBS (shaking at 50 RPM).
- Block with 1× PBS containing 1% LFDM for 30 minutes at room temperature (shake at 50 RPM).
- Rinse with 70 ml of PBS.
- Add your primary antibodies and incubate overnight in the fridge (3–5°C).
- Wash the membrane 5 times (5 minutes each) with PBST containing 0.1% Tween-20 (use 100 ml, shake at 50 RPM).
- Add the Adaptor-Secondary of your choice at 1:5000 dilution and incubate with shake for 30 minutes.
- Dip in DI water (or 1x PBS).
- Incubate with your secondary antibody for example, HRP-Rb anti-Ch (diluted 1:20,000). Use 5 ml per membrane (6×8 cm). Incubate for 30 minutes to 1 hour at room temperature, shaking at 50 RPM.
- Wash 4 times (5 minutes each) with 100 ml of PBST, shaking at 50 RPM.
- Wash 6 more times with 100 ml of PBS, shaking at 50 RPM.
- Add ECL substrate.
- Scan the blot on a C-Digit at high resolution (12 minutes).
- Cut the issue at 5 µm
- Drain and Dry
- Dewax and Dehydrate
- HIER using Thermo's PT Module - HIER buffer: Tris-EDTA pH 9.00
- Stain the slides: Buffer wash - Peroxide Block (5 min) - Buffer wash (2x) - Protein Block (5 min) - Buffer wash (2x) - Primary antibody (30 min) - Buffer wash (2x) - Adaptor-Secondary - Buffer wash - HRP-Polymer (30 min) - Buffer wash (3x) - Water wash (1x)
- Add DAB
- Counterstain
- Coverslip
- Restrictions
- For Research Use only
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- Format
- Lyophilized
- Reconstitution
- Reconstitute in 500 µL DI water.
- Buffer
- Contains Osenses-developed Signal Enhancer and Background Quencher. Preservative: 0.02% benzalkonium chloride
- Preservative
- Benzalkonium chloride
- Storage
- 4 °C,-20 °C
- Storage Comment
- Maintain the lyophilised/reconstituted antibodies frozen at -20°C for long term storage and refrigerated at 2-8°C for a shorter term.
- Expiry Date
- 12 months
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- Target
- IgG
- Abstract
- IgG Products
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