DYKDDDDK (binds to Sigma FLAG®, clone FG4R)

Name: DYKDDDDK (binds to Sigma FLAG®, clone FG4R)
Brand: IKP
SKU: AS15 2871
Price: 259 EUR
Availability: InStock

AS15 2871 | Clonality: monoclonal | Host: Mouse | Reactivity: N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins

DYKDDDDK (binds to Sigma FLAG®, clone FG4R) in the group Tag Antibodies / DYKDDDDK (binds to SigmaFLAG®) at Agrisera AB (Antibodies for research) (AS15 2871)

Product Information

Immunogen

KLH-conjugated DYKDDDDK (FLAG) synthetic peptide

Host Mouse

Clonality Monoclonal

Subclass/isotype IgG1

Purity Total IgG. Protein A purified in 10mM PBS, pH 7.2, with 1% BSA and 0.05% sodium azide.

Format Liquid

Quantity 50 µg

Storage Store at -20°C for up to 2 years.

Tested applications Dot blot (Dot), ELISA (ELISA), Immunohistochemistry (IHC), Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 1 : 1000-5000 (1-5ng) (WB) (incubate for one hour at room temperature), 1 : 500-1 :2000 (IS) For best results with other assays (Dot, ELISA, IP), please determine optimal working dilution by end user

Reactivity

Confirmed reactivity N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples application example

western blot using anti-FLAG antibody

20 µg of total protein from Chlamydomonas reinhardtii Flag; 2 (1) 1xFlag 5’+ICL (1x FLAG coding sequence : GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (2); 1xFlag 5’+ICL (1x FLAG codingsequence :GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (3); ICL+ 3xFlag 3’end (3xFLAG codingsequence : GAC TAC AAG GAC CAC GAC GGT GAC TAC AAG GAC CAC GAC ATC GAC TAC AAG GAC GACGACGAC AAG TGA.) (4); 1xFlag 5’+ICL+ 3xFlag 3’end (5) were separated on 12 % SDS-PAGE and blotted ON (30V) to PVDF using tank transfer. Blots were blocked with 5 % non-fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-mouse IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with BM ECL kit according to the manufacturer's instructions. Exposure time was seconds.

Courtesy of Dr. Nadine Coosemans, Remacle lab, University of Liège, Belgium

Additional information

Additional information Working dilution for Dot Blot, ELISA and IP needs to be determined experimentally.

Antibody is purified on Protein A.

Background

Background FLAG-tag is a tag that can be added to a protein sequence motif DYKXXD. It can be used for affinity chromatography, to separate recombinant protein from wt proteins. It can also be used to isolate protein complexes with multiple subunits.

Product citations

immunogen:	KLH-conjugated DYKDDDDK (FLAG) synthetic peptide
Sub class:	IgG1
Host:	Mouse
Clonality:	Monoclonal
Purity:	Total IgG. Protein A purified in 10mM PBS, pH 7.2, with 1% BSA and 0.05% sodium azide.
Format:	Liquid
Quantity:	50 µg
storage:	Store at -20°C for up to 2 years.
tested applications:	Dot blot (Dot), ELISA (ELISA), Immunohistochemistry (IHC), Immunoprecipitation (IP), Western blot (WB)
recommended dilution:	1 : 1000-5000 (1-5ng) (WB) (incubate for one hour at room temperature), 1 : 500-1 :2000 (IS) For best results with other assays (Dot, ELISA, IP), please determine optimal working dilution by end user
Confirmed reactivity:	N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	application example 20 µg of total protein from Chlamydomonas reinhardtii Flag; 2 (1) 1xFlag 5’+ICL (1x FLAG coding sequence : GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (2); 1xFlag 5’+ICL (1x FLAG codingsequence :GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (3); ICL+ 3xFlag 3’end (3xFLAG codingsequence : GAC TAC AAG GAC CAC GAC GGT GAC TAC AAG GAC CAC GAC ATC GAC TAC AAG GAC GACGACGAC AAG TGA.) (4); 1xFlag 5’+ICL+ 3xFlag 3’end (5) were separated on 12 % SDS-PAGE and blotted ON (30V) to PVDF using tank transfer. Blots were blocked with 5 % non-fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-mouse IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with BM ECL kit according to the manufacturer's instructions. Exposure time was seconds. Courtesy of Dr. Nadine Coosemans, Remacle lab, University of Liège, Belgium
additional information:	Working dilution for Dot Blot, ELISA and IP needs to be determined experimentally. Antibody is purified on Protein A.
background:	FLAG-tag is a tag that can be added to a protein sequence motif DYKXXD. It can be used for affinity chromatography, to separate recombinant protein from wt proteins. It can also be used to isolate protein complexes with multiple subunits.