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DYKDDDDK 标记 抗体

ABIN1112984 产品详细信息, 供应商: Log in to see
抗原
适用
标记
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宿主
小鼠
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标记
非结合性
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应用范围
Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
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Supplier
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Supplier Product No.
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Available images

'独立验证'标志
Lot Number L21/2013G
Method validated Immunofluorescence
Positive Control HEK293 cells transfected with DYKDDDDK Tag
Negative Control Untransfected HEK293 cells
Notes Signal was detected in positive control samples but not in negative control samples.
Primary Antibody
  • Antibody: DYKDDDDK Tag antibody
  • Catalog number: ABIN1112984
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  • Supplier catalog number: Log in to see
  • Lot number: L21/2013G
Secondary Antibody
  • Antibody: Goat anti-mouse IgG (H+L) IF488 conjugated antibody
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  • Supplier catalog number: Log in to see
  • Lot number: S6/2013S
Controls
  • Positive control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged protein (specimen
  • known to contain the target protein).
  • Negative control: Untransfected HEK293 cells from ATCC (specimen known to not contain the target
  • protein).
  • Isotype control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged protein treated with
  • isotype mouse IgG control instead of the primary antibody. Any staining observed is due to non-specific
  • binding of secondary antibody.
  • Secondary antibody only control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged
  • protein treated with Goat anti-Mouse IF488 secondary antibody only. Any staining observed is due to
  • non-specific binding of secondary antibody.
实验流程
  • HEK293 cells were grown directly on coverslips and transfected with a DYKDDDDK tagged protein.
  • Cells were fixed with 4% paraformaldehyde in PBS for 15 min at room temperature (RT).
  • Fixed cells were rinsed three times in PBS for 5 min each at RT.
  • Cells were blocked in 1 X PBS / 5% normal goat serum / 0.3% Triton X-100 for 20 min at RT.
  • Cells were incubated with primary antibody diluted 1:100 in Universal Antibody Dilution Buffer (Electron
  • Microscopy Sciences, 25885-05) overnight at 4°C.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Cells were incubated with secondary antibody diluted 1:150 in Universal Antibody Dilution Buffer
  • (Electron Microscopy Sciences, 25885-05) for 60 min at RT in dark.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Coverslips were mounted on slides with Fluoro-Gel II with DAPI (Electron Microscopy Sciences Cat. No: 17985-50).
  • Stained cells were imaged with an EIDAQ 100 fluorescent microscope.
Experimental Notes None
Validation Images
Immunofluorescence validation image for anti-DYKDDDDK Tag antibody (ABIN1112984) Figure 1: : Immunofluorescence analysis of cultured cells using DYKDDDDK Tag antibody...
免疫原 A synthetic peptide DYKDDDDK conjugated to KLH.
序列 DYKDDDDK
亚型 IgG1
别名 Flag Tag
物质类 Tag
背景 DYKDDDDK-Tag, or DYKDDDDK octapeptide, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. The peptide sequence of the DYKDDDDK-Tag is: N-DYKDDDDK-C (1012 Da). It can be used in conjunction with other affinity tags, for example a polyhistidine tag (His-tag), HA-tag or myc-tag. It can be fused to the C-terminus or the N-terminus of a protein.
应用备注 WB (1:1000-10000), IP (1:100-500), IF (1:200-1000). Other applications have not been tested. The optimal dilutions should be determined by end user.
板类型 Strips (12 x 8)
限制 仅限研究用
状态 Liquid
缓冲液 PBS (pH7.4), 50% Glycerol and 0.02% Sodium Azide.
储存液 Sodium azide, Thimerosal (Merthiolate)
储存条件 4 °C
储存方法 Store at 2-8 °C for 1 month. Or aliquot and store at -20 °C or below for long term. Avoid repeated freeze and thaw cycles.