没错,这是一类专为消除二抗检测时的交叉反应和本底背景的抗体!

Thermo Scientific™ Superclonal™ 超级二抗代表了重组抗体技术的突破,可在多种应用中准确且灵敏的检测小鼠、兔和山羊种属一抗,其采用专门的筛选和生产过程,得到特异性的重组山羊或兔的二抗混合物,可与单克隆抗体表位准确结合,且同时具有多克隆抗体的多表位覆盖性(如H+L)和高灵敏度。每一种Superclonal超级二抗经过专门的制备和优化,有助于取得绝佳的ELISA、Western blot和细胞成像结果。

订购信息

“Superclonal results were better than expected. Specifically, the Superclonal did not exhibit any background or noise whatsoever. Really bright and vibrant resolution on the staining. I was confident that the signals were specific to the targets and not the antibody background.”

—Nathan Schurman, Research Diabetes Technician


Superclonal secondary antibodies help enable precise and accurate detection

Invitrogen Superclonal secondary antibodies are designed to accurate and precise results. The endogenous tubulin of U-87 MG (human neuroblastoma) cells were labeled with anti-alpha tubulin primary antibody, which was then detected with Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate antibody (green). Simultaneously, DISC1 was labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 555 conjugate (red). Cell nuclei (blue) were stained with DAPI (blue).


Features of Superclonal secondary antibodies

  • Developed as recombinant monoclonal antibodies to enable precise and accurate detection
  • Formulated to recognize both heavy- and light-chain epitopes (H+L) of target IgG molecules
  • Selected and optimized for use with cell imaging, ELISA, and western blotting applications
  • Offered in four types: goat anti-mouse (GAM), goat anti-rabbit (GAR), rabbit anti-mouse (RAM), rabbit anti-goat (RAG)
  • Available unconjugated and conjugated with biotin, horseradish peroxidase (HRP), and selected Invitrogen™ Alexa Fluor™ dyes
 
 

Lot-to-lot consistency

Representative images demonstrating lot-to-lot consistency of performance of Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate in immunocytochemistry. No primary control to assess background is shown. Endogenous HIF-1 alpha in U20S human osteocarcinoma cells was labeled (green) with Rabbit anti-HIF-1 alpha primary antibody, and visualized using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate (0.4 µg/mL, 1:2500). Nuclei (blue) were stained with Invitrogen™ SlowFade™ Gold Antifade Mountant using DAPI (1:50). F-actin (red) was stained with Alexa Fluor 594 Phalloidin (1:200). 

Superclonal超级二抗特点:

  • 专为消除一抗检测中的交叉反应性而研发
  • 重组的单克隆抗体可实现最准确且灵敏的检测
  • 可识别靶标IgG分子的重链和轻链表位 (H+L)
  • 经过专门的筛选和优化,可用于细胞成像、ELISA和Western blot
  • 可提供4种种属类型:山羊抗小鼠 (GAM)、山羊抗兔 (GAR)、兔抗小鼠 (RAM)、兔抗山羊 (RAG)
  • 可选择多种标记形式:生物素、辣根过氧化物酶 (HRP) 和Alexa Fluor™染料标记及无标记等。

Compare the technology

Superclonal secondary antibodies compared to traditional secondary antibodies

  Polyclonal antibodies
(pAb)
Superclonal secondary antibodies Monoclonal antibodies (mAb)
How they are made Affinity purify (by positive and/or negative selection) antibodies from the serum of immunized animals Produce and characterize numerous individual AOF recombinant antibodies, then carefully screen, select, and pool specific ones Isolate and select a desired antibody-producing cell from host immunized animal, then culture it to produce a single antibody clone
What they are A large, undefined pool of antibodies from the host serum, selected by affinity purification but otherwise having unknowable sets of specific epitope and binding-affinity characteristics Precisely characterized sets of specific AOF recombinant antibodies having known, complementary sets of epitopes and affinity binding features Well characterized single antibody clones having one specific epitope and affinity binding features.
Epitope coverage and signal amplification

Excellent

Broad epitope coverage for the target IgG (e.g., heavy and light chain, H+L) ensures good sensitivity and signal amplification for the target primary antibody

Excellent

Selected and pooled to provide complementary, specific and optimal epitope coverage (e.g., H+L) and binding quality to maximize performance in key applications

Poor

Binds to a single epitope Can select for a clone that has no off-target binding (i.e., background or cross-reactivity)

Specificity and lot-to-lot consistency

Poor

  • Affinity purification (e.g., pre-adsorption) from the source serum provides only limited specificity (background and cross-reactivity are common limitations of pAbs)
  • Animal variability and purification process can result in an unknown amount of lot variability

Excellent

  • Clones selected and combined in sets designed to eliminate off-target binding (i.e., cross-reactivity) in the tested applications
  • Recombinant technology helps ensure that every lot comprises exactly the same clones

Very good

  • Can select for a clone that has no off-target binding
  • Propagation of monoclonal cell line can result in clonal drift; sequence of antibody is not definitively known
Host and target species available
  • Goat anti-mouse (GAM)
  • Goat anti-rabbit (GAR)
  • Rabbit anti-mouse (RAM)
  • Rabbit anti-goat (RAG)
  • Many others
  • Goat anti-mouse (GAM)
  • Goat anti-rabbit (GAR)
  • Rabbit anti-mouse (RAM)
  • Rabbit anti-goat (RAG)

Traditional monoclonal antibody production uses mouse or rat as the host. Monoclonals are seldom used as secondary antibodies.

Superclonal超级二抗可消除交叉反应,背景极低

使用抗核干因子一抗标记HeLa细胞核,随后使用相应的Alexa Fluor™ 488二抗(绿色)检测:(A) 高度交叉吸附的山羊抗小鼠IgG (H+L) 二抗,或 (B) 山羊抗小鼠IgG (H+L) SuperClonal超级二抗。细胞核采用DAPI(蓝色)染色,肌动蛋白微丝使用Alexa Fluor 594 Phalloidin(红色)标记。发现使用Superclonal超级二抗检测,细胞质染色显著减少,说明其特异性更强。

Superclonal超级二抗与传统二抗的比较

  多克隆抗体
(pAb)
Superclonal™ 超级二抗 单克隆抗体 (mAb)
制备方法 从免疫动物的血清中得到的亲和纯化(通过阳性和/或阴性选择)抗体 生产和表征大量单个重组抗体后,仔细筛选、选择并对特异性抗体进行富集 从宿主免疫动物中分离和筛选可产生目的抗体的细胞,培养细胞产生一种抗体克隆
成分 宿主血清中经亲和纯化选择的大量不明确的抗体,具有不可知的特异性表位簇和结合亲和性的特性。 精确表征的特异性重组抗体集合,具有已知的表位的互补集簇和亲和结合的特性 经过良好表征的单克隆抗体,具有一种特异性表位和亲和结合的特性
表位覆盖和信号放大

极佳

对目标IgG(如,重链和轻链,H+L)的广泛表位覆盖,保证了对目标一抗高灵敏度和信号放大

极佳

通过选择和富集,可产生具有互补性和特异性的理想表位覆盖(如,H+L)和结合能力,使其在常见应用中的性能最大化

较差

与单一表位结合,可选择出无脱靶结合(如,背景或交叉反应)的克隆

特异性和批次一致性

较差

  • 对血清的亲和纯化(如,预吸附)只能产生有限的特异性(背景和交叉反应是多克隆抗体的共同缺陷)
  • 动物差异和纯化过程可导致某些未知的批次差异

极佳

  • 被选定然后混合的重组克隆可消除检测应用中的脱靶结合(即,交叉反应)
  • 重组技术有助于确保每一批次均包含相同的克隆

很好

  • 可选择出无脱靶结合的克隆
  • 单克隆细胞系的增殖可导致克隆漂移;无法明确抗体的序列
宿主和靶向种属
  • 山羊抗小鼠 (GAM)
  • 山羊抗兔 (GAR)
  • 兔抗小鼠 (RAM)
  • 兔抗山羊 (RAG)
  • 其他种属
  • 山羊抗小鼠 (GAM)
  • 山羊抗兔 (GAR)
  • 兔抗小鼠 (RAM)
  • 兔抗山羊 (RAG)

传统单克隆抗体的生产使用小鼠或大鼠作为宿主。单克隆很少作为二抗使用。

Superclonal超级二抗可消除交叉反应,背景极低

使用抗核干因子一抗标记HeLa细胞核,随后使用相应的Alexa Fluor™ 488二抗(绿色)检测:(A) 高度交叉吸附的山羊抗小鼠IgG (H+L) 二抗,或 (B) 山羊抗小鼠IgG (H+L) SuperClonal超级二抗。细胞核采用DAPI(蓝色)染色,肌动蛋白微丝使用Alexa Fluor 594 Phalloidin(红色)标记。发现使用Superclonal超级二抗检测,细胞质染色显著减少,说明其特异性更强。

Superclonal超级二抗与传统二抗的比较

  多克隆抗体
(pAb)
Superclonal™ 超级二抗 单克隆抗体 (mAb)
制备方法 从免疫动物的血清中得到的亲和纯化(通过阳性和/或阴性选择)抗体 生产和表征大量单个重组抗体后,仔细筛选、选择并对特异性抗体进行富集 从宿主免疫动物中分离和筛选可产生目的抗体的细胞,培养细胞产生一种抗体克隆
成分 宿主血清中经亲和纯化选择的大量不明确的抗体,具有不可知的特异性表位簇和结合亲和性的特性。 精确表征的特异性重组抗体集合,具有已知的表位的互补集簇和亲和结合的特性 经过良好表征的单克隆抗体,具有一种特异性表位和亲和结合的特性
表位覆盖和信号放大

极佳

对目标IgG(如,重链和轻链,H+L)的广泛表位覆盖,保证了对目标一抗高灵敏度和信号放大

极佳

通过选择和富集,可产生具有互补性和特异性的理想表位覆盖(如,H+L)和结合能力,使其在常见应用中的性能最大化

较差

与单一表位结合,可选择出无脱靶结合(如,背景或交叉反应)的克隆

特异性和批次一致性

较差

  • 对血清的亲和纯化(如,预吸附)只能产生有限的特异性(背景和交叉反应是多克隆抗体的共同缺陷)
  • 动物差异和纯化过程可导致某些未知的批次差异

极佳

  • 被选定然后混合的重组克隆可消除检测应用中的脱靶结合(即,交叉反应)
  • 重组技术有助于确保每一批次均包含相同的克隆

很好

  • 可选择出无脱靶结合的克隆
  • 单克隆细胞系的增殖可导致克隆漂移;无法明确抗体的序列
宿主和靶向种属
  • 山羊抗小鼠 (GAM)
  • 山羊抗兔 (GAR)
  • 兔抗小鼠 (RAM)
  • 兔抗山羊 (RAG)
  • 其他种属
  • 山羊抗小鼠 (GAM)
  • 山羊抗兔 (GAR)
  • 兔抗小鼠 (RAM)
  • 兔抗山羊 (RAG)

传统单克隆抗体的生产使用小鼠或大鼠作为宿主。单克隆很少作为二抗使用。