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Cat. No.: AVAG-0.5 (for 0.5ml)

Cat. No.: AVAG-2 (for 2ml)

Cat. No.: AVAG-10 (for 10ml)

Description

Anti-V5 Affinity Gel, also known as Anti-V5 IP Gel, Anti-V5 immunoprecipitation gel, or Anti-V5 agarose gel, is formed by covalently coupling high-quality monoclonal mouse-derived V5 antibodies to agarose. It can specifically bind to proteins containing the V5 tag in common protein expression systems such as mammalian cells, bacteria, or yeast, enabling immunoprecipitation (IP) or purification of fusion proteins or protein complexes with the V5 tag.

V5 tag, Flag tag, Myc tag, HA tag, His tag, and GST tag are some of the most common tags found on expression vectors. Fusion expression with these tags allows for convenient detection of the target protein and proteins that interact with the target protein. It also facilitates easy purification of the target protein. The V5 tag is a peptide consisting of 14 amino acid residues (GKPIPNPLLGLDST) derived from the monkey parainfluenza virus V5. By genetically fusing the nucleotide sequence encoding the V5 tag to the 5' or 3' end of the target gene using recombinant techniques, the target protein with the V5 tag can be expressed. The V5 tag offers the following advantages: it typically does not interact with the target protein and, in most cases, does not affect the function of the target protein. As a protein tag, the V5 tag allows for subsequent detection, localization, and functional analysis of the target gene using V5 antibodies (AF2894), Anti-V5 magnetic beads (P2141), or Anti-V5 affinity gels (P2289). Based on these advantages, the V5 tag has been widely used in various aspects of protein expression, purification, identification, protein-protein interactions, and functional studies.

Anti-V5 Affinity Gel, also known as Anti-GKPIPNPLLGLDST Affinity Gel/Beads/Resin or Anti-GKPIPNPLLGLDST IP Gel/Beads/Resin, can specifically bind to fusion proteins with the V5 tag. It is widely used in immunoprecipitation or purification of fusion proteins or protein complexes with the V5 tag.

Features

• This product boasts a high binding capacity and strong specificity for fusion proteins: Each ml of settled gel contains approximately 7.5 mg of V5 antibody, capable of binding about 0.5 mg of fusion protein, with minimal non-specific protein binding.
• Versatile binding to various forms of V5-tagged proteins: This product can specifically bind N-terminal V5 fusion proteins (V5-Protein) and C-terminal V5 fusion proteins (Protein-V5).
• Multiple elution methods available: Depending on the protein's structural integrity, biological function, and subsequent applications, various elution methods can be used, including V5 peptide, acidic solutions, and SDS-PAGE loading buffer. Notably, elution with V5 peptide does not include antibody light and heavy chains, effectively addressing interference in Western blot assays following immunoprecipitation.
• Reusable and cost-effective: Under normal conditions, this product can be reused 3-5 times for purifying the same protein. However, it is not recommended for reuse in immunoprecipitation assays detecting protein-protein interactions.

Parameters

• Product Content: 50% settled gel in 50% glycerol with 10mM phosphate-buffered saline and preservative (pH 7.4)
• Matrix: 4% agarose
• Average Bead Size: ~90 μm
• Antibody: Mouse monoclonal antibody against V5-tag
• Isotype: IgG1
• Molecular Weight of Antibody: Approximately 150 kDa
• Antibody Concentration: Approximately 7.5 mg of V5 antibody per ml of settled gel
• Binding Capacity: Approximately 0.5 mg of V5-tagged protein per ml of settled gel
• Elution Methods: Acidic, alkaline, neutral, peptide competitive, or SDS-PAGE loading buffer elution. Note: When eluted with SDS-PAGE loading buffer, the light (~25 kDa) and heavy (~50 kDa) chains of the antibody will be denatured and released from the gel.
• Reagent Compatibility: Chaotropic reagents will denature the target V5-tagged protein. Do not exceed 0.3M GuHCl or 1.5M urea.
• Application: Suitable for IP, Co-IP, and protein purification.
• Storage: -20°C

Storage

Store at -20°C, valid for one year.

Precautions

• Ensure thorough resuspension before use by inverting several times to mix well.
• The product contains trace amounts of preservative, which should not affect routine protein or protein complex purification and immunoprecipitation. However, if enzyme activity assays are involved, wash the gel with TBS or another suitable solution three times before use to remove any potential interference from the preservative.
• When performing immunoprecipitation or purification, it is recommended to include positive and negative control groups.
• After collecting protein samples, proceed with purification promptly and keep them at 4°C or in an ice bath to minimize protein degradation or denaturation. To effectively inhibit protein degradation, add an appropriate amount of protease inhibitor mix to the protein samples.
• High concentrations of DTT, β-mercaptoethanol, guanidine hydrochloride, etc., may affect the binding of this product to tagged proteins.
• If centrifugation does not completely remove insoluble matter from the protein samples, filter the sample solution with a 0.45 μm filter membrane.
• This product is for scientific research use by trained professionals only and is not intended for clinical diagnosis or treatment, food, or drug use. Do not store in residential areas.
• For your safety and health, please wear a lab coat and disposable gloves while handling.

Related:

• Anti-Flag Affinity Gel v2
• Anti-Flag Affinity Gel
• Anti-Myc Affinity Gel
• Anti-HA Affinity Gel
• Anti-V5 Affinity Gel

Only for research and not intended for treatment of humans or animals

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