DYKDDDDK Tag 抗体
(1 validation)Quick Overview for DYKDDDDK Tag 抗体 (ABIN559688)
抗原
适用
宿主
克隆类型
标记
应用范围
克隆位点
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序列
- DYKDDDDK
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特异性
- Recognizes the N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
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灵敏度
- WB (with ECL): 1-5ng of DYKDDDDK-tagged fusion proteins
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纯化方法
- Protein A affinity chromatography from mouse ascites fluid
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免疫原
- DYKDDDDK (DYKDDDDK) synthetic peptide conjugated to KLH
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亚型
- IgG1
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anti-DYKDDDDK Tag antibody
WB, ELISA, IP 宿主: 兔 Polyclonal unconjugated
anti-DYKDDDDK Tag antibodyWB, ELISA, IF, FACS 宿主: 兔 Polyclonal unconjugated
anti-DYKDDDDK Tag antibodyWB, ELISA, IP 宿主: 小鼠 Monoclonal 5A8E5 unconjugated
anti-DYKDDDDK Tag antibody适用: 标记 WB, ELISA, IP, IHC, FACS, ICC 宿主: 小鼠 Monoclonal 2F11G1 unconjugated
anti-DYKDDDDK Tag antibodyWB, IP, IF, IC 宿主: 小鼠 Monoclonal unconjugated
anti-DYKDDDDK Tag antibody适用: 人 WB 宿主: 兔 Polyclonal RB0493 unconjugated
anti-DYKDDDDK Tag antibody (HRP)WB, ELISA, IF 宿主: 小鼠 Monoclonal 5A8E5 HRP
anti-DYKDDDDK Tag antibodyWB, ELISA, IHC, FACS 宿主: 小鼠 Monoclonal 29E4-G7 unconjugated
anti-DYKDDDDK Tag antibodyWB, IP, IF 宿主: 小鼠 Monoclonal F-tag-01 unconjugated
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应用备注
- WB (with ECL): 1:1000-3000 dilution (incubate for one hour at room temperature) / IS: 1:500-2000 dilution / For best results with other assays (e.g.: Dot, ELISA, IP, etc), please determine optimal working dilution by titration test
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限制
- 仅限研究用
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- by
- Martinelli lab, Neuroscience Department, UConn Health
- No.
- #102796
- 日期
- 2018.02.13
- 抗原
- DYKDDDDK-tag
- Lot Number
- 02017
- Method validated
- Immunofluorescence
- Positive Control
- African green monkey kidney fibroblast-like COS7 cells transfected with BAI3, a transmembrane protein with a surface presented DYKDDDDK-tag
- Negative Control
- untransfected COS7 cells
- Notes
Passed. Signal was observed as expected. There was never any signal on untransfected cells, demonstrating specificity of the antibody.
- Primary Antibody
- ABIN1715174
- Secondary Antibody
- goat anti-mouse AF488 conjugated antibody (Life Technologies, A11029, lot 1829920)
- Full Protocol
- Grow COS7 cells in DMEM (Biological Industries, 01-052-1A, lot 1728619) supplemented with FBS (Hyclone, SH3039602) without antibiotics, at 37°C and 5% CO2 in 1ml on glass coverslips on 24-well plate.
- Transfect cells with calcium phosphate method with a eukaryotic expression construct to express a DYKDDDDK-tagged BAI3, which will be surface presented.
- Grow cells for 24h.
- Fix cells on coverslips in 4% PFA for 15min at 4°C.
- Wash cells 3x for 5min with PBS.
- Block non-specific binding with 5% goat serum in PBS for 1h at RT.
- Incubate cells with primary mouse anti-DYKDDDDK-tag antibody (antibodies-online, ABIN1715174, lot 02017) diluted 1:2000 in blocking solution ON at 4°C.
- Wash cells 3x for 5min with PBS.
- Incubate cells with secondary goat anti-mouse AF488 conjugated antibody (Life Technologies, A11029, lot 1829920) diluted 1:400 in blocking buffer for 1h at RT protected from light.
- Wash cells 3x for 5min with PBS.
- Mount coverslips on glass slides in Fluoromount-G with DAPI mounting medium (ThermoFisher Scientific, 00-4959-52, lot B2215-N915).
- Image acquisition on a Zeiss Axiovert epifluorescence with with a 40x objective, using a Zeiss ApoTome for optical sectioning.
- Experimental Notes
生效 #102796 (Immunofluorescence)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![COS7 cells were transfected to express a DYKDDDDK-tagged and surface presented BAI3 protein. Indirect immunoflouresence using ABIN1715174 reveals specific signal for the DYKDDDDK-tag (middle) on the cell surface. No colocalization of the ABIN1715174 and nuclear stain (DAPI, left) was observed (merge, right). No signal was ever observed in any untransfected cell.]()
COS7 cells were transfected to express a DYKDDDDK-tagged and surface presented BAI3 protein. Indirect immunoflouresence using ABIN1715174 reveals specific signal for the DYKDDDDK-tag (middle) on the cell surface. No colocalization of the ABIN1715174 and nuclear stain (DAPI, left) was observed (merge, right). No signal was ever observed in any untransfected cell.
Full Methods -
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浓度
- 1mg/mL
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缓冲液
- 10mM PBS, pH 7.2, 0.05% NaN3
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储存液
- Sodium azide
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注意事项
- This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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储存条件
- -20 °C
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- DYKDDDDK Tag
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别名
- FLAG Tag
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物质类
- Tag
抗原
-
