MOU-IP Anti-mouse IgG (HRP) for IP/CoIP

产品名称：MOU-IP Anti-mouse IgG (HRP) for IP/CoIP

货号： IF9204

品牌：Engibody

简介：IP专用抗小鼠二抗， CoIP专用抗小鼠二抗 ，用于下游WB实验中，消除免疫沉淀抗体的轻重链干扰

MOU-IP™ Anti-mouse IgG for IP (HRP Conjugated) secondary antibody is western blotting reagents that enable the trouble-free detection of western blotted target protein bands from upstream IP/CoIP, without interference from denatured IgG (IP primary antibody). This allows to detect the (co-)immunoprecipitated protein without interfering by the IgG heavy (50 kDa) and light chains (25 kDa). In general, this interference tends to originate from traditional secondary antibodies which recognize primary antibodies released with the antigen during the immunoprecipitation procedure or endogenous IgGs from the lysate itself.

MOU-IP™ Anti-mouse IgG for IP (HRP Conjugated) only recognize native (non-reduced) primary antibodies (from mouse) and therefore the bands of heavy and light chains is highly minimized, if the immunoprecipitated antigen-antibody complex is fully reduced and denatured before downstream WB.

Description

MOU-IP™ Anti-mouse IgG (native conformation antibody specific) for IP (HRP Conjugated)

Reactivity

Mouse

Tested applications

WB : 1/1000-1/2000. Optimal dilutions should be determined by the end user.

Specificity

This secondary antibody is specific to native mouse polyclonal/monoclonal antibody

Immunogen

The antibody was developed in goat using the nature mouse IgG as the immunogen and then develop monoclonal antibody.

Clonality

Monoclonal, clone number: 7G9

Isotype

Goat IgG

Form

The antibody in 10 mM PBS, pH 7.4, 50% glycerol with 10 mg/mL (1% w/v) BSA as a stabilizer and 0.01% (w/v) thimerosal as preservative.

Storage instruction

Store at -20°C, Avoid freeze / thaw cycle. Do not aliquot the antibody.

Conjugation

Horseradish Peroxidase (HRP)

Host

Goat

MOU-IP™ IP专用的抗小鼠IgG二抗（天然构象完整分子抗体特异性）（HRP偶联）是一种蛋白IP-WB试剂，其能够无障碍地检测来自上游IP/CoIP的蛋白质印迹目标蛋白带，而不受变性IgG的干扰（IP一抗）。这允许在不受IgG重链（50kDa）和轻链（25kDa）干扰的情况下检测（共）免疫沉淀蛋白。通常，这种干扰倾向于源于传统的二抗，其识别在免疫沉淀过程中与抗原一起释放的一抗或来自裂解物本身的内源性IgG的轻链和重链。

MOU-IP™ 如果免疫沉淀的抗原-抗体复合物在下游WB之前被还原和变性，则用于IP的抗小鼠IgG（HRP标记）仅识别天然（非还原）一抗（来自小鼠），因此重链和轻链的条带不会显示。

描述

MOU-IP™ IP专用的抗小鼠IgG二抗（天然构象完整分子抗体特异性）（HRP偶联）

反应性

识别小鼠完整抗体分子

经过测试的应用实验

WB:1/1000-1/2000。最佳稀释度应由最终用户确定。

特异性

该二抗对天然小鼠多克隆/单克隆抗体完整分子具有特异性识别，同时不识别小鼠抗体单独轻重链

免疫原

以天然小鼠IgG为免疫原，在山羊身上制备抗体，然后制备只识别小鼠抗体完整分子同时不识别小鼠抗体单独轻重链的单克隆抗体。

克隆性

单克隆，克隆号：7G9

同型对照

山羊IgG

类型

抗体在10 mM PBS中，pH 7.4，50%甘油，10 mg/mL（1%w/v）BSA作为稳定剂，0.01%（w/v）硫柳汞作为防腐剂。

存储说明

储存于-20°C，避免冷冻/解冻循环。不要分装抗体。

偶联标记

辣根过氧化物酶（HRP）

抗体来源宿主

山羊

Figure 1. MOU-IP™ Anti-mouse IgG for IP (HRP) secondary antibody images: Western blotting.

IP sample preparation:

target protein (PCNA) was immunoprecipitated from 0.5 mL cell lysate of 1x107 Jurkat cells with 5 µg anti-human PCNA mouse monoclonal antibody and protein A/G agarose beads (Engibody, IF0001).

WB conditions:

the immunoprecipitated antigen-antibody complex should be boiled for 5-10 minutes in SDS sample buffer with a increase in SDS amount if required, make sure that the complex is fully reduced and denatured before downstream WB.

then electrophoresis, transferred to a PVDF membrane, and incubate with an anti- PCNA mouse monoclonal antibody.

Secondary Antibody Detection:

Lane 1: Detection with MOU-IP™ Anti-mouse IgG for IP (HRP) (CAT: IF9204)

The heavy and light chains can not be seen, confirming that although the immunoprecipitating heavy and light-chains are present, detects only native antibody and not denatured heavy and light-chains.

Lane 2: Detection with a traditional goat anti-mouse IgG (H&L) (HRP) secondary antibody (CAT: AT0098)

Figure 2. A schematic representation of IP-WB workflow by using MOU-IP™ secondary antibody.

Using primary antibody coupled beads

MOU-IP™ Anti-mouse IgG for IP (HRP Conjugated) only recognize native (non-reduced) primary antibodies (from mouse) and therefore the bands of heavy chain and light chain are highly minimized, if the immunoprecipitated antigen-antibody complex is fully reduced and denatured before downstream WB.

Figure 3. A schematic representation of IP-WB workflow by using MOU-IP™ secondary antibody.

Using protein A/G Beads + primary antibody

MOU-IP™ Anti-mouse IgG for IP (HRP Conjugated) only recognize native (non-reduced) primary antibodies (from mouse) and therefore the bands of heavy chain and light chain are highly minimized, if the immunoprecipitated antigen-antibody complex is fully reduced and denatured before downstream WB.