GFP标签小鼠单克隆抗体(3D3),琼脂糖偶联

Name: GFP标签小鼠单克隆抗体(3D3),琼脂糖偶联
Brand: Abbkine
SKU: ABT2023
Price: 1658 CNY
Availability: InStock

Agarose Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)

产品货号

ABT2023

产品特点：

琼脂糖微球已预偶联1 mg高纯度小鼠抗GFP单克隆抗体，浓度≥1 mg Ab/mL settled resin，开盖即用。

小鼠IgG1亚型，高特异性识别GFP及常见突变体，背景结合低，信噪比优异。

兼容哺乳动物细胞、细菌等多种表达体系，适用于IP、Co-IP及pull-down实验。

提供PBS保存体系（含0.02% NaN₃及50% PBS），4 °C可稳定保存一年，避免反复冻融。

选择规格

1 mL

¥1658

现货（次日发货）

5 mL

¥5658

现货（次日发货）

🎉 限时优惠

买2送1活动进行中！购买任意2个规格产品，免费赠送100μL装一支。

活动截止时间：2024年1月31日

产品概述

亚科因生物研发的Agarose Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)是一款将高亲和力小鼠单克隆抗体与琼脂糖树脂共价偶联的免疫沉淀工具，可直接用于捕获带GFP标签的重组蛋白，无需额外添加Protein A/G，实现“一步法”IP富集。

绿色荧光蛋白（GFP）源自维多利亚水母，由238个氨基酸组成，在蓝光或紫外光激发下发出509 nm绿色荧光，已成为分子与细胞生物学中最常用的报告标签之一。本抗体（克隆号3D3）特异性识别GFP及其常见突变体（如EGFP、sfGFP），通过亲和层析从小鼠腹水中纯化后，与琼脂糖微球共价偶联，形成稳定的固相载体。当含有GFP标签的样品与琼脂糖-抗体复合物孵育时，目标蛋白被选择性捕获；经洗涤去除非特异结合后，可直接用于Western blot、质谱或活性分析，显著缩短实验流程。

应用领域

广泛用于分子互作研究、信号通路验证、重组蛋白富集与质谱鉴定、GFP融合蛋白表达水平监测等场景，尤其适合IP实验流程的简化与高通量需求。

产品参数

中文名称	GFP标签小鼠单克隆抗体(3D3),琼脂糖偶联
英文名称	Agarose Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)
产品货号	ABT2023
免疫原	重组蛋白
宿主	小鼠
反应性	哺乳动物#细菌
标签	GFP
检测类型	IP
克隆类型	单克隆
亚型	小鼠IgG1
纯化工艺	通过使用特异性免疫原的亲和层析，从小鼠腹水中纯化抗体。
偶联物	琼脂糖树脂
产品形式	液体形式
储存缓冲液	50% gel slurry suspended in PBS, pH 7.4, containing 0.02% Sodium Azide.
保存建议	从发货之日起，4°C可稳定保存一年。避免反复冻融或者涡旋。
运输条件	冰袋运输（蓝冰）
警告	本文列出的产品仅供研究使用，不适用于人类或临床诊断。我们产品所推荐应用，不是建议使用我们的产品去违反任何专利或许可证。对于使用本产品可能发生的专利侵权或其他违规行为，我们不承担任何责任。

使用说明

Reagent Required but Not Supplied

Elution Buffer: 0.1 M Glycine-HCl pH 3.0.

Elution Buffer:

Neutralization Buffer: 1 M Tris-HCl, pH 8.5.

Neutralization Buffer:

Assay Procedure

A. Preparation of Agarose

Note: Per 500 μL of protein sample add 20 μL (total 40 μL suspension) Agarose.

Note:

Add Agarose to a 1.5 mL centrifuge tube. Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.
Add 1 mL 1×TBS to re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3 times. Add 20 μL 1×TBS to re-suspend Agarose.

Add Agarose to a 1.5 mL centrifuge tube. Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.

Add 1 mL 1×TBS to re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3 times. Add 20 μL 1×TBS to re-suspend Agarose.

B. Immunoprecipitation

Add 500 μL protein samples to the processed Agarose, and incubate at room temperature for 1-2 h or overnight at 4℃ (It is recommended to use vertical rotating mixer with Low-speed rotation).
Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.
Add 1 mL 1×TBS, and re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3-5 times, until OD280 of the supernatant is lesser than 0.05.
Elution
1. Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add 100 μL (5 times volume of Agarose) 1×SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100℃ for 5 min, then Centrifuge at 800 rpm for 1 min, and collect the supernatant to a new tube for SDS-PAGE and Western Blotting analysis.
2. Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add 100 μL (5 times volume of Agarose) Elution Buffer to the tube and mix well, incubate at room temperature for 5-10 min (It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4℃, and collect the supernatant which is GFP-Tag protein and its complex to a new tube, and immediately add 10 μL Neutralization Buffer to adjust the pH to 7.0-8.0. In order to improve the elution efficiency, elution can be repeated, and combine the same samples. Place GFP-Tag protein and its complex on ice to be used, or store at -20 ℃ /-80 ℃ for long-term. It is recommended to add 100 μL 1×SDS-PAGE Loading Buffer to Agarose precipitation to test the effect of immunoprecipitation and elution.

Add 500 μL protein samples to the processed Agarose, and incubate at room temperature for 1-2 h or overnight at 4℃ (It is recommended to use vertical rotating mixer with Low-speed rotation).

Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.

Add 1 mL 1×TBS, and re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3-5 times, until OD280 of the supernatant is lesser than 0.05.

Elution

Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add 100 μL (5 times volume of Agarose) 1×SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100℃ for 5 min, then Centrifuge at 800 rpm for 1 min, and collect the supernatant to a new tube for SDS-PAGE and Western Blotting analysis.
Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add 100 μL (5 times volume of Agarose) Elution Buffer to the tube and mix well, incubate at room temperature for 5-10 min (It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4℃, and collect the supernatant which is GFP-Tag protein and its complex to a new tube, and immediately add 10 μL Neutralization Buffer to adjust the pH to 7.0-8.0. In order to improve the elution efficiency, elution can be repeated, and combine the same samples. Place GFP-Tag protein and its complex on ice to be used, or store at -20 ℃ /-80 ℃ for long-term. It is recommended to add 100 μL 1×SDS-PAGE Loading Buffer to Agarose precipitation to test the effect of immunoprecipitation and elution.

Elution

Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add 100 μL (5 times volume of Agarose) 1×SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100℃ for 5 min, then Centrifuge at 800 rpm for 1 min, and collect the supernatant to a new tube for SDS-PAGE and Western Blotting analysis.
Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add 100 μL (5 times volume of Agarose) Elution Buffer to the tube and mix well, incubate at room temperature for 5-10 min (It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4℃, and collect the supernatant which is GFP-Tag protein and its complex to a new tube, and immediately add 10 μL Neutralization Buffer to adjust the pH to 7.0-8.0. In order to improve the elution efficiency, elution can be repeated, and combine the same samples. Place GFP-Tag protein and its complex on ice to be used, or store at -20 ℃ /-80 ℃ for long-term. It is recommended to add 100 μL 1×SDS-PAGE Loading Buffer to Agarose precipitation to test the effect of immunoprecipitation and elution.

Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add 100 μL (5 times volume of Agarose) 1×SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100℃ for 5 min, then Centrifuge at 800 rpm for 1 min, and collect the supernatant to a new tube for SDS-PAGE and Western Blotting analysis.

Denatured elution:

Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add 100 μL (5 times volume of Agarose) Elution Buffer to the tube and mix well, incubate at room temperature for 5-10 min (It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4℃, and collect the supernatant which is GFP-Tag protein and its complex to a new tube, and immediately add 10 μL Neutralization Buffer to adjust the pH to 7.0-8.0. In order to improve the elution efficiency, elution can be repeated, and combine the same samples. Place GFP-Tag protein and its complex on ice to be used, or store at -20 ℃ /-80 ℃ for long-term. It is recommended to add 100 μL 1×SDS-PAGE Loading Buffer to Agarose precipitation to test the effect of immunoprecipitation and elution.

Acid elution:

Note: a) For a few samples, due to differences in target proteins, the binding of GFP-Tag and Anti-GFP antibody is very strong, and the effect of Acid elution may be poor. Therefore, SDS-PAGE Loading Buffer denaturation elution method is recommended as a priority; b) Due to the difference of target protein, the elution efficiency of acid elution method also varies to some extent. If the requirement of elution efficiency is high, the pH value of acidic eluent can be adjusted appropriately between 2.5-3.1, and the pH value or quantity of corresponding neutralizing solution should be adjusted appropriately. For example, 100 μL Acid Elution Buffer (0.1 M Glycine-HCl, pH 2.8) and 15 μL Neutralizing Buffer (1 M Tris-HCl, pH 8.5).

Note:

Recommended Products

Catalog No.	Product Name
ABT2013	Agarose Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)
ABT2043	Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)
ABT2053	Agarose Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)
ABT2063	Agarose Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)
ABT2173	Agarose Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5)

Catalog No.Product NameCatalog No.Product NameABT2013Agarose Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)ABT2013Agarose Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)ABT2043Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)ABT2043Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)ABT2053Agarose Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)ABT2053Agarose Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)ABT2063Agarose Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)ABT2063Agarose Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)ABT2173Agarose Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5)ABT2173Agarose Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5)

Disclaimer

The reagent is only used in the field of scientific research, not suitable for clinical diagnosis or other purposes.

常见问题

Q: 普通荧光二抗能不能做WB呢？应该如何成像呢？

A: 荧光western blotting的结果需要特殊的仪器检测，即一种荧光成像仪器。即运用荧光成像仪可以对荧光WB的实验结果进行成像和数据采集分析。如果希望应用荧光成像做WB，建议选用Abbkine DyLight 680和DyLight 800荧光二抗，货号分别为A23710、A23720、A23910、A23920。

Q: Abbkine品牌的二抗的浓度为多少呢？

A: 0.5 mg/ml。

Q: 抗体中甘油的作用是什么？

A: 甘油是可以减慢抗体分子之间的碰撞，起到抗体保护剂的作用。

文献引用

Lipid droplet accumulation in Wdr45-deficient cells caused by impairment of chaperone-mediated autophagic degradation of Fasn.

杂志名称: Lipids in Health and Disease | 作者: Xiong, Qiuhong, et al.

IF: 4 | 发表时间: 2024

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