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TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag

Name: TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag
SKU: ABIN6809839
Availability: OutOfStock

IP, WB

Western Blot-Monoclonal Antibody to detect conjugated proteins Monoclonal Antibody to detect conjugated proteins detects both C terminal linked and N terminal linked tagged recombinant proteins by western blot.

Mouse IP / Western Blot: Caspase 7 was immunoprecipitated from 0.

Immunoprecipitation of VHH-EGFR-FLAG single chain antibody.

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产品编号 ABIN6809839

发货至: 中国

Datasheet as PDF

应用范围

Immunoprecipitation (IP), Western Blotting (WB)

原理

This TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag allows for the detection of FLAG®-tagged recombinant protein present in cell lysates

品牌

TrueBlot®

特异性

This TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag allows for the detection of FLAG®-tagged recombinant protein present in cell lysates provided by the user. Mouse TrueBlot® ULTRA Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against Anti-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.

产品特性

The FLAG® epitope is a small but highly immunogenic peptide DYKDDDDK (N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C) which allows fusion proteins to retain their original conformation and function. The hydrophilic character of this sequence increases the likelihood that it will be located on the surface of FLAG®-tagged recombinant proteins for reaction with anti-DYKDDDDK (FLAG®) antibodies. The Anti-DYKDDDDK (FLAG®) antibody included in this Western blot kit detects FLAG®-tagged recombinant proteins fused with FLAG® tag either at the amino-terminus or carboxy-terminus of the protein. The TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples containing the DYKDDDDK (FLAG®) Epitope Tag using TrueBlot monoclonal secondary antibody in conjunction with Rockland's Antibody for the detection of FLAG™ conjugated proteins (MOUSE) Monoclonal Antibody (see application notes). Mouse IgG TrueBlot® ULTRA is the unique horseradish peroxidase conjugated Anti-Mouse IgG imonoclonal secondary antibody which enables detection of immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your conventional HRP Anti-Mouse IgG immunoblotting secondary antibody. It is easy to generate publication-quality IP/WB data with Mouse IgG TrueBlot® ULTRA. Mouse IgG TrueBlot ULTRA is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of mouse IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse IgG TrueBlot ULTRA with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/immunoblot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

组件
TrueBlot® IP and WB Kit for 6X HIS Epitope Tag Components:
1. Mouse IgG TrueBlot ULTRA: 50 μL ABIN1589976
2. TrueBlot Enhancer Solution: 25 mL
3. TrueBlot Blocker: 10 g
4. TrueBlot Assay Buffer: 30 mL 20X
5. TrueBlot Substrate A: 12.5 mL
6. TrueBlot Substrate B: 12.5 mL
7. Anti-Mouse Ig IP Beads: 2.5 mL Binds 0.4 mg Ig/mL beads
8. Anti-6X HIS (MOUSE) Monoclonal Antibody: 100 µg
9. Western Blot Incubation Tray.
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应用备注

Western Blot : 1:1000
ImmunoPrecipitation: 1-10 μg / 10^7 cells/1 mL lysate

说明
Mouse IgG TrueBlot® ULTRA is provided as 1000X solution. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). Mouse IgG TrueBlot® ULTRA is an HRP-conjugated monoclonal secondary antibody reacting with mouse IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments.
Note that there are three key procedural considerations:
1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands.
2. Immunoprecipitate should be completely reduced.
3. BLOTTO/Milk should be used as the blocking protein for the immunoblot.
Special Notes: Upon initial use of the IP beads, we recommend that the vial be inverted several times to get the beads into suspension. We recommend using a large bore pipet to pipet up the liquid for use. For storage of the opened vial of beads, we recommend that the vial cap be sealed with parafilm to help prevent evaporation of the buffer. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user.
限制

仅限研究用
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Buffer: 0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Stabilizer: 0.1 mg/mL Bovine Serum Albumin (BSA) - IgG and Protease free, 50 % (v/v) Glycerol
Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity!

储存条件

4 °C

储存方法

Store Kit at 2-8 °C, except Mouse TrueBlot® ULTRA and Antibody for the detection of FLAG™ conjugated proteins (MOUSE) Monoclonal Antibody, which should be stored at -20 °C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.