抗体名: flag标签抗体偶联agarose beads(affinity gel) 抗体英文名: anti flag affinity gel 靶点: flag标签融合蛋白 浓度: 50% slurry 应用范围: Immunoprecipitation (IP), 40 ul of gel suspension (20μl of packed beads) for ~1000μg of crude total protein solution. Optimal dilutions/concentrations should be determined by the end user. 宿主: 小鼠 适应物种: all species 标记物: agarose beads 克隆性: 单克隆 保存条件: –20 °C 形态: 液体 亚型: mouse IgG1 免疫原: This monoclonal antibody is produced by immunizing mouses with a synthetic peptide (KLH-coupled) containing DYKDDDDK. 规格: 1ml
Reagent | Effect | Notes |
Chaotropic agents (e.g., urea, guanidine HCl) | Denatures the immobilized anti FLAG antibody | Do not use any reagent that contains these types of components since it will denature the anti FLAG antibody on the gel and destroy its ability to bind the FLAG fusion proteins. Low concentrations of urea (1 M or less) can be used. |
Reducing agents (such as DTT, DTE, 2-mercaptoethanol) | Reduces the disulfide bridges holding the anti FLAG antibody chains together | Do not use any reagent that contains these types of components since it will reduce the disulfide linkages in the anti FLAG antibody on the gel and destroy its ability to bind the FLAG fusion proteins. |
Sodium dodecyl sulfate (SDS) | Denatures the immobilized anti FLAG antibody | Do not use any reagent that contains this detergent in the lysis and washing buffers since it will denature the anti FLAG antibody on the gel and destroy its ability to bind the FLAG fusion proteins. It is included in the sample loading buffer for removal of protein for immunoprecipitation, but the gel cannot be reused. |
Deoxycholate | Interferes with anti FLAG antibody binding to FLAG proteins | Do not use any reagent that contains this detergent since it will inhibit the anti FLAG antibody from binding to FLAG fusion proteins. |
Reagent | Effect | Notes |
TWEEN 20, 5% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 5%, but do not exceed. |
TRITON X-100 5% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 5%, but do not exceed. |
IGEPAL CA-630, 0.1% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.1%, but do not exceed. |
CHAPS, 0.1% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.1%, but do not exceed. |
Digitonin, 0.2% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.2%, but do not exceed. |
Sodium chloride, 1.0 M or less | Reduces nonspecific protein binding to the gel by reducing ionic interactions | May be used up to recommended concentration of 1.0 M, but do not exceed. |
0.1 M glycine HCl, pH 3.5 | Elutes FLAG protein from the gel | Do not leave the beads in glycine HCl for longer than 20 minutes. Longer incubation times will begin to denature the anti FLAG antibody |
Problem | Possible Cause | Solution |
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