AM20780AG-N DYKDDDDK Epitope Tag antibody

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0.5 ml / €340.00
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Quick Overview

Mouse anti All Species DYKDDDDK Epitope Tag 3B9


Product Description for DYKDDDDK Epitope Tag

Mouse anti All Species DYKDDDDK Epitope Tag 3B9.
Presentation: Agarose
Product is tested for Immunoprecipitation.

Properties for DYKDDDDK Epitope Tag

Product Category Primary Antibodies
Quantity 0.5 ml
Synonyms D-tag, ECS Epitope Tag, ECS-tag, FLAG Epitope Tag, FLAG-tag
Presentation Agarose
Reactivity All Species
Applications IP
Clonality Monoclonal
Clone 3B9
Host Mouse
Isotype IgG2b
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Synthetic peptide containing epitope DYKDDDDK (KLH-coupled).
Application Immunoprecipitation.
Background Anti-DYKDDDDK-Tag Mouse mAb (Agarose Conjugated) is a monoclonal anti-DYKDDDDK antibody covalently linked to agarose; the agarose enables immunoprecipitation (IP) of DYKDDDDK tagged proteins or co-immunoprecipitation (Co-IP) of their interacting partners.
Protocols Immunoprecipitation procedure
The work can be performed in 1.5 ml micro-centrifuge tubes or in spin columns.
1. Thoroughly resuspend the Anti-DYKDDDDK Agarose by inverting the tube or vial several times.

2. Add 20-50 μl 50% slurry of Anti-DYKDDDDK Agarose into cell lysate using a wide-bore pipette tip. Note: the lysate should be fresh, and for a well expressed tagged protein, 200 μl lysate (200-500 g total protein) usually yields a good IP result.

3. Incubate with gentle mixing for 2 h to overnight at 4°C.

4. Wash the beads with 1 ml TBS buffer or lysis buffer, such as RIPA (50 mM Tris HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate), centrifuge for 3 min at 2,000× g, and discard the supernatant. Wash 3 times, avoid losing beads during washes.

5. Elution of the DYKDDDDK tagged protein.
Option 1. Elution with elution buffer.
Add 30-50 μl elution buffer to the beads, gently tap the tube to mix well, immediately centrifuge for 3 min, transfer the supernatant very carefully to a fresh tube (Avoid transferring any beads).
Note: Neutralize the eluant immediately by add 1 μl of 1.5 M Tris, pH 9.0 per 20 μl Elution buffer.

Option 2. Elution with DYKDDDDK peptide
Add 30-50 μl DYKDDDDK peptide solution (100 g/ml DYKDDDDK peptide in TBS buffer), gently tap the tube to mix well, incubate for 10 min, centrifuge for 3 min, and transfer the supernant to a fresh tube. TBS buffer: 50 mM Tris HCl, 150 mM NaCl, pH 7.4.

Option 3. Elution with SDS loading buffer
Add 30 μl 2 SDS loading buffer, gently tap the tube to mix well, boil at 100°C for 5 min, centrifuge for 3 min, transfer the supernatant to a fresh tube.
Note: in this case, the supernatant contains not only the binding proteins, but also IgG (heavy and light chains).

6. Prepare SDS-PAGE gel for western blotting or proceed to other assays
Storage Store the antibody undiluted at 2-8°C.
Shelf life: one year from despatch.
Buffer System:
50% slurry in storage buffer (1× PBS, pH 7.4, containing 0.09% sodium azide).
Recommended elution buffer: 0.2 M Glycine, pH 2.5
Liquid purified Ig fraction
This antibody reacts to DYKDDDDK Epitope Tag.

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