DYKDDDDK tag polyclonal antibody
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More Files
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Western Blot (Recombinant protein)
DYKDDDDK tag polyclonal antibody (Cat # PAB10340) to detect FLAG™ conjugated proteins detects both C terminal linked and N terminal linked FLAG™ tagged recombinant proteins by western blot.
This antibody was used at a dilution of 1 : 2,500 to detect 1.0 ug of recombinant protein containing either the FLAG™ epitope tag linked at the carboxy (C) or the amino (N) terminus of the recombinant protein.
A 4-20% gradient gel was used to resolve the protein by SDS-PAGE.
The protein was transferred to nitrocellulose using standard methods.
After blocking, the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1 : 10,000 dilution of IRDye® 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 30 min at room temperature.
LICOR's Odyssey® Infrared Imaging System was used to scan and process the image.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptides of DYKDDDDK tag.
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to DYKDDDDK tag.
Host
Rabbit
Specificity
This antibody reacts with FLAG conjugated proteins.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:90000-1:250000)
Western Blot (1:2000-1:10000)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM potassium phosphate buffer, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Recombinant protein)
DYKDDDDK tag polyclonal antibody (Cat # PAB10340) to detect FLAG™ conjugated proteins detects both C terminal linked and N terminal linked FLAG™ tagged recombinant proteins by western blot.
This antibody was used at a dilution of 1 : 2,500 to detect 1.0 ug of recombinant protein containing either the FLAG™ epitope tag linked at the carboxy (C) or the amino (N) terminus of the recombinant protein.
A 4-20% gradient gel was used to resolve the protein by SDS-PAGE.
The protein was transferred to nitrocellulose using standard methods.
After blocking, the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1 : 10,000 dilution of IRDye® 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 30 min at room temperature.
LICOR's Odyssey® Infrared Imaging System was used to scan and process the image.Enzyme-linked Immunoabsorbent Assay
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Publication Reference
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Identification of new tag sequences with differential and selective recognition properties for the anti-FLAG monoclonal antibodies M1, M2 and M5.
Slootstra JW, Kuperus D, Plückthun A, Meloen RH.
Molecular Diversity 1997 Mar; 2(3):156.
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Double tagging recombinant A1- and A2A-adenosine receptors with hexahistidine and the FLAG epitope. Development of an efficient generic protein purification procedure.
Robeva AS, Woodard R, Luthin DR, Taylor HE, Linden J.
Biochemical Pharmacology 1996 Feb; 51(4):545.
Application：IP, Hamster, CHO-K1 cells.
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Vectors for expression and secretion of FLAG epitope-tagged proteins in mammalian cells.
Chubet RG, Brizzard BL.
Biotechniques 1996 Jan; 20(1):136.
Application：ICC, IHC, IP, WB-Tr, Monkey, COS-1 cells.
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Identification of new tag sequences with differential and selective recognition properties for the anti-FLAG monoclonal antibodies M1, M2 and M5.
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