TrueBlot® Anti-Rabbit Ig IP Beads

Catalog No. ABIN1589964

Supplier Product No.: 00-8800-25

Product Details

Reactivity: Rabbit

Host: Goat

Clonality: Polyclonal

Conjugate: Agarose Beads

Application: Immunoprecipitation (IP), Western Blotting (WB)

Supplier Product No.: 00-8800-25

Supplier: Rockland

Brand: TrueBlot®

Characteristics: TrueBlot® anti-Rabbit Ig IP Beads are a suspension of activated agarose beads coupled with goat anti-rabbit IgG. It is suitable for precipitation of rabbit IgGs used as the primary antibodies in immunoprecipitation assays. The beads are in suspension and will settle upon storage. Prior to use, mix the vial gently (do not vortex) to ensure delivery of proper bead volume.
Conjugation Name: Agarose beads for TrueBlot®

Components: TrueBlot® Anti-Rabbit Ig IP Beads

Application Details

Application Notes: Immunoprecipitation Dilution: TrueBlot® anti-Rabbit Ig IP Beads (binds 2.5 mg Ig/mL beads) have been reported for use in IP
Western Blot Dilution: Use with Rabbit TrueBlot® (ABIN1589974)

Comment:

Upon initial use of this product, we recommend that the vial be inverted several times to get the beads into suspension. We recommend to use a large bore pipet to pipet up the liquid for use. For storage of the opened vial, we recommend that the vial cap be sealed with parafilm to help prevent evaporation of the buffer.

Assay Procedure:

Preparation of Immunoprecipitated Sample for SDS-PAGE:
1. Preclear cell lysate: Add 50 µL of Anti-Rabbit IgG Beads and 500µL of cell lysate sample to a microcentrifuge tube and incubate on ice for
3. minutes. Spin at 10,000xg for 3 minutes and transfer thesupernatant to a new microcentrifuge tube.
2. Immunoprecipitation: Add 5 µg of primary antibody to the microcentrifuge tube containing the precleared lysate. Incubate on ice for 1 hour. Add 50 µL of Anti-Rabbit IgG Beads. Incubate for 1 hour on a rocking platform. Spin the microcentrifuge tube at 10000xg for 1 minute Remove supernatant completely and wash the (pelleted) beads 3 times with 500 µL of Lysis Buffer (50mM Tris HCl pH 8.0, 150mM NaCl,1 % NP-40).
3. Prepare sample for SDS-PAGE: After the last wash, aspirate supernatant, and add 100 µL Laemmli Buffer (with 50 mM DTT or 1 %ß-mercaptoethanol, final) to bead pellet. Vortex and heat to 90-100 °C for
1. minutes. Spin at 10000xg for 3 minutes, collect supernatant, and load onto the gel. Avoid loading Anti-Rabbit Ig Beads.
Note: The supernatant can be stored at -20 °C for future use. After thawing, add fresh dithiothreitol and heat as above. Centrifuge the sample at 10000xg for 1 minute in a microcentrifuge tube to pellet any Anti-Rabbit Ig Beads and immediately transfer an aliquot of the supernatant to gel wells.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer:

Buffer: 0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2

0.01 % (w/v) Sodium Azide

Stabilizer: None

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Do not freeze.
Sensitive to light.
Prior to use, mix the vial gently (do not vortex) to ensure delivery of proper bead volume.

Storage: 4 °C

Storage Comment: Store vial at 4 °C prior to opening.

Expiry Date: 6 months

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