DYKDDDDK Epitope Tag Antibody

MAB8529 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

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MAB8529

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Detection of DYKDDDDK (FLAG®) Epitope Tag by Western Blot.

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DYKDDDDK Epitope Tag Antibody Summary

Specificity

Detects N-terminal and C-terminal DYKDDDDK (FLAG^®) epitope tagged proteins in Western blot.

Source

Recombinant Monoclonal Rabbit IgG Clone # 1042E

Purification

Protein A or G purified from cell culture supernatant

Immunogen

DYKDDDDK epitope tag

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

See below

Simple Western

1 µg/mL

See below

Flow Cytometry

0.25 µg/10⁶ cells

See below

Immunoprecipitation

1-4 µg/100 µg cell lysate

HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1 and N-terminal DYKDDDDK-tagged RNF20, see our available Western blot detection antibodies.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot

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Detection of DYKDDDDK (FLAG^®) Epitope Tag by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with
C-terminal DYKDDDDK-tagged Abro1,
N-terminal DYKDDDDK-tagged UCHL5, and N-terminal DYKDDDDK-tagged RNF20. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody
(Catalog # MAB8529) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for DYKDDDDK-tagged proteins at approximately 55 kDa, 40 kDa, 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry

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Detection of DYKDDDDK (FLAG^®) in HEK293 Human Cell Line Transfected with DYKDDDDK-tagged Proteins by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with DYKDDDDK-tagged proteins was stained with Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody (Catalog # MAB8529, filled histogram) or isotype control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111).

Simple Western

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Detection of DYKDDDDK (FLAG^®) Epitope Tag by Simple Western^TM. Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1 and N-terminal DYKDDDDK-tagged RNF20, loaded at 0.1 mg/mL. Specific band were detected for DYKDDDDK-tagged proteins at approximately 64 and 136 kDa (as indicated) using 1 µg/mL of Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody
(Catalog # MAB8529). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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