As a result of the conjugation, there may be some M2 antibody that is not conjugated to the resin, but is associated with the resin and may appear in acid elutions as heavy and light chain when using the anti-mouse IgG conjugated secondary antibody. We recommend a acid wash (0.1 M glycine-HCL pH 3.5) and neutralization of the resin (do not allow the acid wash to sit on the resin longer than 20 minutes) prior to applying the lysate. Another way to avoid this is to use a directly conjugated FLAG antibody for detection such as product A8592 ant-FLAG M2 HRP, or the rabbit anti-FLAG polyclonal antibody, F7425.
추천 제품
클론
M2, monoclonal
분석물 화학적 분류
proteins
기술
affinity chromatography: suitable
immunoprecipitation (IP): suitable
Matrix
4% agarose bead; 45-165μm bead size
동형
IgG1
용량
≥0.6 mg/mL, gel binding capacity
배송 상태
wet ice
저장 온도
−20°C
유사한 제품을 찾으십니까? 방문 제품 비교 안내
일반 설명
EZ view Red Anti-FLAG M2 Affinity Gel is a resin that consists of Anti-FLAG M2 antibody, covalently bonded to 4% Agarose beads. The affinity gel is used to bind FLAG fusion proteins to samples, such as cell lysates and tissue, for purification of FLAG-tagged proteins in preparation of immunoprecipitation assays. Red dye enhances visability for more efficient results. Agarose beads bind at N-terminal, Met-N-terminal and C-terminal FLAG fusion proteins, 3xFLAG-tagged fusion proteins.
특이성
Suitable for purifying N-terminal, Met-N-terminal, C-terminal FLAG fusion proteins, 3xFLAG fusion proteins.
애플리케이션
Immunoprecipitation (IP) of FLAG- and 3xFLAG-tagged fusion proteins.
Elution - FLAG peptide, Glycine, pH 3.5, 3x FLAG peptide
Learn more product details in our FLAG® application portal.
Elution - FLAG peptide, Glycine, pH 3.5, 3x FLAG peptide
Learn more product details in our FLAG® application portal.
물리적 형태
1:1 (v/v) suspension in PBS containing 50% glycerol and 15 ppm Kathon
법적 정보
ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
EZview is a trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
면책조항
FLAG® Affinity Gels, FLAG® tag, 3xFLAG®tag, DYKDDDDK tag
관련 제품
제품 번호
설명
가격
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
가장 최신 버전 중 하나를 선택하세요:
시험 성적서(COA)
Lot/Batch Number
이미 열람한 고객
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프로토콜
Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels
관련 콘텐츠
Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.
이온교환, 크기 배제 및 단백질 친화성 크로마토그래피를 포함한 방법을 사용하는 재조합형 단백질 정제를 위한 단백질 정제 기법, 시약 및 프로토콜.
단백질 발현 기술은 연구, 치료제, 백신 생산을 지원하는 다양한 발현 시스템에 사용됩니다.
Protein expression technologies for various expression systems supporting research, therapeutics, and vaccine production.
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, I see bands at 20-25 kDa and 50-60 kDa appearing in my Westerns that are not my FLAG-tagged protein. How can I prevent this?
1 답변-
도움이 되었습니까?
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What is the binding capacity of the Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, resin?
1 답변-
The binding capacity of the resin must be ? 0.6 mg/mL to meet specifications. This capacity will vary from lot to lot.
도움이 되었습니까?
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, I have a lot of non-specific proteins that are eluting with my FLAG-tagged protein. How can I get rid of these?
1 답변-
One way to remove non-specific proteins is to pre-bind the protein lysate with unconjugated resin. We recommend product 4B200 for this purpose. Other methods would be to increase the stringency of the washes by increasing salt concentration (the resin can tolerate up to 1M NaCl) or including detergents that are compatible with the resin.
도움이 되었습니까?
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What is the Department of Transportation shipping information for this product?
1 답변-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
도움이 되었습니까?
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, how can I elute my protein?
1 답변-
Elution with the peptide is the most gentle method. Acid elution (0.1 M glycine-HCL pH 3.5) is a more stringent method of elution, and should be evaluated for its effect on your protein if it is to be used in downstream applications. Boiling the resin in sample buffer is the most denaturing condition. If this condition is used, the resin cannot be re-used, due to the presence of SDS and/or reducing agents.
도움이 되었습니까?
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?
1 답변-
If you have a 3X FLAG-tagged protein, then you will need to use the 3X FLAG peptide. If you have a 1X FLAG-tagged protein, you can use the 1X FLAG peptide or the 3X FLAG peptide. We have not noticed a significant difference in elution efficiency by using a 3X FLAG peptide on a 1X FLAG-tagged protein.
도움이 되었습니까?
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