DYKDDDDK Tag anticorps

N° du produit ABIN1112984

L’anticorps Souris anti-DYKDDDDK Tag a été validé pour WB, IP et IF et a été validé indépendamment. Il convient pour détecter DYKDDDDK Tag dans des échantillons de .

Aperçu rapide pour DYKDDDDK Tag anticorps (ABIN1112984)

Antigène: DYKDDDDK Tag

Reactivité: Veuillez nous consulter SVP

Hôte: Souris

Conjugué: Cet anticorp DYKDDDDK Tag est non-conjugé

Application: Western Blotting (WB), Immunoprecipitation (IP), Immunofluorescence (IF)

Détail du produit anti-DYKDDDDK Tag anticorps

Séquence: DYKDDDDK

Immunogène: A synthetic peptide DYKDDDDK conjugated to KLH.

Isotype: IgG1

Information d'application

Indications d'application: 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.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #028889 (Immunofluorescence)

by: Reveal Biosciences

No.: #028889

Date: 07.11.2013

Numéro du lot: L21/2013G

Application validée: Immunofluorescence

Contrôle positif: HEK293 cells transfected with DYKDDDDK Tag

Contrôle négative: Untransfected HEK293 cells

Conclusion: Signal was detected in positive control samples but not in negative control samples.

Validation Images

Figure 1: : Immunofluorescence analysis of cultured cells using DYKDDDDK Tag antibody (Catalog number ABIN1112984, Lot number L21/2013G). Flag is present in the positive control (HEK293 cells transfected with DYKDDDDK Tag panel 1, transfected cells are indicated by green fluorescent staining) and absent from the negative control (untransfected HEK293 cells panel 2). No staining was observed in the isotype (panel 3) and secondary only (panel 4) controls. DAPI was used as a nuclear counterstain (indicated by blue fluorescent staining). Scale bar = 25 µm.

Protocole

Anticorps primaire:

• Antibody: DYKDDDDK Tag antibody
• Catalog number: ABIN1112984
• Lot number: L21/2013G

Anticorps secondaire:

• Antibody: Goat anti-mouse IgG (H+L) IF488 conjugated antibody
• Lot number: S6/2013S

Full Protocol:

• 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.

Notes: None

Stockage

Format: Liquid

Buffer: PBS (pH7.4), 50% Glycerol and 0.02% Sodium Azide.

Agent conservateur: Sodium azide, Thimerosal (Merthiolate)

Stock: 4 °C

Stockage commentaire: 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.

Détails sur DYKDDDDK Tag

Antigène: DYKDDDDK Tag

Autre désignation: Flag Tag

Classe de substances: Tag

Sujet: 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.