DYKDDDDK Tag anticorps

Détails pour le produit réf. ABIN559688
Antigène
Reactivité
Tag
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32
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3
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1
1
Hôte
Souris
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139
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11
Clonalité (Clone)
Monoclonal ()
Conjugué
Inconjugué
32
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6
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2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
Application
Dot Blot (DB), Immunostaining (ISt), Immunoprecipitation (IP), ELISA, Western Blotting (WB)
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119
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Options
'Independent Validation' signe
Antigène DYKDDDDK-tag
Numéro du lot 02017
Application validée Immunofluorescence
Contrôle positif African green monkey kidney fibroblast-like COS7 cells transfected with BAI3, a transmembrane protein with a surface presented DYKDDDDK-tag
Contrôle négative untransfected COS7 cells
Conclusion

Passed. Signal was observed as expected. There was never any signal on untransfected cells, demonstrating specificity of the antibody.

Anticorps primaire ABIN1715174
Anticorps secondaire goat anti-mouse AF488 conjugated antibody (Life Technologies, A11029, lot 1829920)
Protocole
  • 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.
Validation Images
Immunofluorescence validation image for anti-DYKDDDDK Tag antibody (ABIN1715174) COS7 cells were transfected to express a DYKDDDDK-tagged and surface presented BAI3 p...
Immunogène DYKDDDDK (DYKDDDDK) synthetic peptide conjugated to KLH
Séquence DYKDDDDK
Clone FG4R
Isotype IgG2b
Specificité Recognizes the N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
Sensibilité WB (with ECL): 1-5ng of DYKDDDDK-tagged fusion proteins
Purification Protein A affinity chromatography from mouse ascites fluid
Antigène
Autre désignation FLAG Tag
Classe de substances Tag
Indications d'application 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
Restrictions For Research Use only
Concentration 1mg/mL
Buffer 10mM PBS, pH 7.2, 0.05% NaN3
Agent conservateur Sodium azide
Précaution d'utilisation This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Stock -20 °C
Images (Fournisseur)
 image for anti-DYKDDDDK Tag antibody (ABIN559688) 1:1000 (1μg/mL) Ab dilution probed against HEK293 cells transfected with DYKDDDDK-tag...
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