Chicken anti-Sheep IgG Antibody

Catalog No. ABIN7595354

Quick Overview for Chicken anti-Sheep IgG Antibody (ABIN7595354)

Target: IgG

Clonality: Polyclonal

Application: ELISA, Immunohistochemistry (IHC), Western Blotting (WB), Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Adaptor Antibody (AAb)

Isotype: IgY

Product Details

Reactivity: Sheep

Host: Chicken

Purpose: Chicken anti sheep IgY. Functions as an adapter to convert a primary antibody from one species into another.

Specificity: Specific for sheep IgG

Purification: Purified

Application Details

Protocol: Western Blot Protocol

1. Run the SDS-PAGE at 250V.
2. Soak the gel for 15 minutes in transfer buffer, then rinse with DI water.
3. Soak the membrane for 5 minutes in DI water.
4. Semi-dry transfer at 10V.
5. After transfer, rinse the membrane once with PBS, then wash 3× for 5 minutes each with PBS (shaking at 50 RPM).
6. Block with 1× PBS containing 1% LFDM for 30 minutes at room temperature (shake at 50 RPM).
7. Rinse with 70 ml of PBS.
8. Add your primary antibodies and incubate overnight in the fridge (3–5°C).
9. Wash the membrane 5 times (5 minutes each) with PBST containing 0.1% Tween-20 (use 100 ml, shake at 50 RPM).
10. Add the Adaptor-Secondary of your choice at 1:5000 dilution and incubate with shake for 30 minutes.
11. Dip in DI water (or 1x PBS).
12. 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.
13. Wash 4 times (5 minutes each) with 100 ml of PBST, shaking at 50 RPM.
14. Wash 6 more times with 100 ml of PBS, shaking at 50 RPM.
15. Add ECL substrate.
16. Scan the blot on a C-Digit at high resolution (12 minutes).

IHC-P Protocol

1. Cut the issue at 5 µm
2. Drain and Dry
3. Dewax and Dehydrate
4. HIER using Thermo's PT Module - HIER buffer: Tris-EDTA pH 9.00
5. 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)
6. Add DAB
7. Counterstain
8. Coverslip

Restrictions: For Research Use only

Handling

Format: Lyophilized

Reconstitution: Reconstitute in 500 µL DI water.

Buffer: Contains 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

Target Details

Target: IgG