DYKDDDDK tag polyclonal antibody (HRP)
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More Files
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Western Blot
Western blot analysis of Lane 1: BAP FLAG C-Term. Lane 2: BAP FLAG N-Term. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: DYKDDDDK tag polyclonal antibody (HRP) (Cat # PAB31810) at 1:1,000 for 60 min at RT. Blocking: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT. Predicted/Observed size: 49 kDa, 49 kDa for BAP FLAG. Other band(s): None.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptides of FLAG epitope tag. The antibody is conjugated with Horseradish Peroxidase (HRP).
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to DYKDDDDK tag.
Sequence
DYKDDDDK
Host
Rabbit
Specificity
Carboxy and amino terminal linked FLAG tagged recombinant proteins. This antibody can be used to identify fusion proteins containing the FLAG epitope. The antibody recognizes the epitope tag fused to the amino- and carboxy- termini of targeted proteins. This affinity purified antibody is directed against the FLAG motif and is useful in determining its presence in various assays. This polyclonal anti-FLAG tag antibody detects over-expressed proteins containing the FLAG epitope tag at either the N or C terminus. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins.
Form
Lyophilized
Conjugation
HRP
Preparation Method
This antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase Cleavage Site (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the carboxy terminal end to facilitate coupling. This antibody reacts with FLAG conjugated proteins.
Purification
Affinity purification
Isotype
IgG
Recommend Usage
ELISA (1:20000)
Immunohistochemistry (1:500-1:2000)
Western Blot (1:2000-1:5000)
The optimal working dilution should be determined by the end user.Storage Buffer
Lyophilized from 0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2 (10 mg/mL BSA (immunoglobulin and protease free), 0.01% (w/v) gentamicin sulfate). Do NOT add sodium azide!
Storage Instruction
Store vial at 4°C prior to restoration.
After reconstitution with 100 uL deionized water (or equivalent), aliquot contents and freeze at -20°C or below for extended storage.
Aliquot to avoid repeated freezing and thawing.
Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. -
Applications
Western Blot
Western blot analysis of Lane 1: BAP FLAG C-Term. Lane 2: BAP FLAG N-Term. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: DYKDDDDK tag polyclonal antibody (HRP) (Cat # PAB31810) at 1:1,000 for 60 min at RT. Blocking: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT. Predicted/Observed size: 49 kDa, 49 kDa for BAP FLAG. Other band(s): None.Immunohistochemistry
Enzyme-linked Immunoabsorbent Assay
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