Kaninchen anti-Karpfen IgM Antikörper

Produktnummer ABIN7595370

Kurzübersicht für Kaninchen anti-Karpfen IgM Antikörper (ABIN7595370)

Target: IgM

Klonalität: Polyklonal

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

Isotyp: IgG

Produktdetails

Reaktivität: Karpfen

Wirt: Kaninchen

Verwendungszweck: Rabbit anti Carp IgM (CH2-4) IgG. Functions as an adapter to convert a primary antibody from one species into another.

Spezifität: Specific for carp IgM constant heavy chains: CH2 CH3 CH4

Aufreinigung: Purified

Anwendungsinformationen

Applikationshinweise: Use at 1:500 dilution.

Protokoll: 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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Lyophilized

Rekonstitution: Reconstitute in 500 µL DI water.

Buffer: Contains Signal Enhancer and Background Quencher. Preservative: 0.02% benzalkonium chloride

Konservierungsmittel: Benzalkonium chloride

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Maintain the lyophilised/reconstituted antibodies frozen at -20°C for long term storage and refrigerated at 2-8°C for a shorter term.

Haltbarkeit: 12 months

Antigendetails

Target: IgM