DYKDDDDK Tag Antikörper
Kurzübersicht für DYKDDDDK Tag Antikörper (ABIN559688)
Target
Reaktivität
Wirt
Klonalität
Konjugat
Applikation
Klon
- 
    - 
                                            Sequenz
- DYKDDDDK
- 
                                            Spezifität
- Recognizes the N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
- 
                                            Sensitivität
- WB (with ECL): 1-5ng of DYKDDDDK-tagged fusion proteins
- 
                                            Aufreinigung
- Protein A affinity chromatography from mouse ascites fluid
- 
                                            Immunogen
- DYKDDDDK (DYKDDDDK) synthetic peptide conjugated to KLH
- 
                                            Isotyp
- IgG1
 
- 
                                            
- 
    
- 
    - 
                                            Applikationshinweise
- 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
- 
                                            Beschränkungen
- Nur für Forschungszwecke einsetzbar
 
- 
                                            
- 
        - by
- Martinelli lab, Neuroscience Department, UConn Health
- No.
- #102796
- Datum
- 13.02.2018
- Antigen
- DYKDDDDK-tag
- Chargennummer
- 02017
- Validierte Anwendung
- Immunofluorescence
- Positivkontrolle
- African green monkey kidney fibroblast-like COS7 cells transfected with BAI3, a transmembrane protein with a surface presented DYKDDDDK-tag
- Negativkontrolle
- untransfected COS7 cells
- Bewertung
- Passed. Signal was observed as expected. There was never any signal on untransfected cells, demonstrating specificity of the antibody. 
- Primärantikörper
- ABIN1715174
- Sekundärantikörper
- 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.
 
- Anmerkungen
 Validierung #102796 (Immunofluorescence)![Erfolgreich validiert 'Independent Validation' Siegel]()  ![Erfolgreich validiert 'Independent Validation' Siegel]() Validierungsbilder Validierungsbilder![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. 
                        
                    Protokoll 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. 
                        
                    Protokoll
- 
    - 
                                            Konzentration
- 1mg/mL
- 
                                            Buffer
- 10mM PBS, pH 7.2, 0.05% NaN3
- 
                                            Konservierungsmittel
- Sodium azide
- 
                                            Vorsichtsmaßnahmen
- This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 
                                            Lagerung
- -20 °C
 
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- 
    - DYKDDDDK Tag
- 
                                            Andere Bezeichnung
- FLAG Tag
- 
                                            Substanzklasse
- Tag
 Target
- 
                    
 
                                                 (1 Validierung)
 (1 Validierung) 
                 
                                     
                                     
                                    