Anti-GFP小鼠单克隆抗体

产品名称：Anti-GFP小鼠单克隆抗体

货号： AT0028

品牌：Engibody

简介：GFP标签抗体，一管搞定全部实验（WB，IHC，ICC，IF及IP/CoIP）

The green-fluorescent protein (GFP) functions as a bioluminescence energy transfer acceptor in the jellyfish Aequorea that maximally absorbs light at 395 nm and has an emission spectrum that peaks at 509 nm. GFP has become a very useful tool as a fusion protein that reports gene expression, traces cell lineage and defines subcellular protein localization.

Description

Mouse monoclonal antibody to GFP

Reactivity

green fluorescent protein (GFP), Enhanced green fluorescent protein (EGFP)

Tested applications

WB    : 1/2000-1/10000.

IHC    : 1/200-1/5000.

ICC/IF    : 1/200-1/2000.

IP/CoIP    : 1/100-1/500.

Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

Specificity

This antibody detects GFP, EGFP -tagged proteins exogenously expressed in cells or E. coli. This antibody does not detect RFP-tagged proteins.

Please note that the GFP, EGFP tags add approximately 27 kDa to the molecular weight of the fusion protein.

The antibody recognizes the GFP, EGFP tag, which is fused to either the amino-terminus or carboxy-terminus of the target protein.

Immunogen

a Green Fluorescent Protein (GFP) recombinant protein.

Clonality

Monoclonal, clone number: 7C5

Isotype

Mouse IgG2a

Form

Liquid, 100 μg at 1mg/mL

Mouse monoclonal antibody in 10 mM phosphate buffered saline (PBS), pH 7.4, 50% glycerol with 1 mg/mL (0.1% w/v) BSA (IgG free, protease free) as a stabilizer and 0.02% (w/v) sodium azide as preservative.

Storage instruction

Store at -20°C, Avoid freeze / thaw cycle.

Host

Mouse

绿色荧光蛋白（GFP）在水母Aequorea中充当生物发光能量转移受体，其最大吸收395nm处的光。GFP作为一种融合蛋白已成为一种非常有用的工具，可报告基因表达、追踪细胞谱系和确定亚细胞蛋白定位。

描述

抗GFP的小鼠单克隆抗体

反应性

增强型绿色荧光蛋白（EGFP）

已测试的应用实验

WB:1/2000-1/10000。

IHC:1/200-1/5000。

ICC/IF:1/2000-1/2000。

IP/CoIP:1/100-1/500。

尚未在其他应用中测试。最佳稀释/浓度应由最终用户确定。

特异性

这种抗体检测细胞或大肠杆菌中外源表达的GFP、EGFP标记蛋白。该抗体不能检测RFP标记的蛋白质。

请注意，GFP、EGFP标签使融合蛋白的分子量增加了约27kDa。

抗体识别GFP，即EGFP标签，融合到目标蛋白的氨基末端或羧基末端。

免疫原

绿色荧光蛋白（GFP）重组蛋白。

克隆性

单克隆，克隆号：7C5

同型对照

小鼠IgG2a

类型

液体，100μg，1mg/mL

小鼠单克隆抗体在10 mM磷酸盐缓冲盐水（PBS）中，pH 7.4，50%甘油，1 mg/mL（0.1%w/v）BSA（不含IgG，不含蛋白酶）作为稳定剂，0.02%（w/v）防腐剂。

存储说明

-20°C下，避免冷冻/解冻循环。

抗体来源宿主

小鼠

Western Blot: GFP Antibody

Lane 1: HEK293T lysate 50µg (transfected pEGFP-N1-POI).

Lane 2: HEK293T lysate 50µg (transfected pEGFP -C1-POI).

Lane 3: Hela lysate 50µg (transfected pEGFP -N1-POI).

Primary antibody: GFP antibody at 1:5000

Observed band size: 48 kDa

Recommended secondary antibody: Goat anti mouse IgG-HRP (Engibody, Catalog number: AT0098)

Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

Immunocytochemistry / Immunofluorescence:

GFP Antibody(AT0028) - GFP-transfected 293T cells.

mCherry antibody (AT0037) labels mCherry tagged protein

Nuclei were counterstained with DAPI.

Immunoprecipitation: Anti-GFP tag Mouse mAb (Engibody, Catalog number:AT0028)

Analysis of 400 µg cell lysate/extract of GFP-fused gene transfected 293T cells

Lane 1: 30 µg lysate/extract of transfected 293T cell

Lane 2:  Immunoprecipitation of GFP-fused protein by 1:500 of GFP tag Mouse mAb

Lane 3: Control with 1:500 of normal mouse IgG

Recommended immunoprecipitation antibody for IP: Anti-GFP tag Mouse mAb (Engibody, Catalog number:AT0028)

Recommended protein A/G beads: Protein A/G Agarose Beads (Engibody, Catalog number: IF0001), or Protein A/G Magnetic Beads (Engibody, Catalog number: IF0002)

Recommended detection antibody for downstream WB: Anti-GFP tag mouse mAb (Engibody, Catalog number:AT0028)

Recommended secondary antibody for downstream WB: MOU-IP Anti-mouse IgG for IP (HRP) (Engibody, Catalog number:AT9204), so that the heavy and light chains from immunoprecipitation antibody can't be seen in WB.