DYKDDDDK (binds to Sigma FLAG®) (polyclonal)

Name: DYKDDDDK (binds to Sigma FLAG®) (polyclonal)
SKU: AS20 4442
Price: 302 EUR
Availability: InStock

AS20 4442 | Clonality: Polyclonal | Host: Rabbit | Reactivity: DYKDDDDK-Tag

DYKDDDDK (binds to Sigma FLAG®) (polyclonal) in the group Tag Antibodies / DYKDDDDK (binds to SigmaFLAG®) at Agrisera AB (Antibodies for research) (AS20 4442)

DYKDDDDK (binds to Sigma FLAG®) (polyclonal)

Product Information

Immunogen KLH-conjugated synthetic peptide: DYKDDDDK (Sigma FLAG®)

Host Rabbit

Clonality Polyclonal

Purity Immunogen affinity purified serum, in PBS pH 7.4.

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl, of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Western blot (WB)

Recommended dilution 1 : 1000 - 1: 5000 (WB)

Expected | apparent MW Depends upon fusion protein partner

Reactivity

Confirmed reactivity DYKDDDDK epitope tag (Sigma FLAG®), fused to proteins in plant cells

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

Application examples

Application examples Western blot using anti-FLAG epitope tag polyclonal anmtibodies

Recombinant FLAG-protein was expressed in bacteria and purified with affinity chromatography. 1.5 µg of FLAG protein was used per in vitro SUMOylation assay in two different conditions: line A. in 30°C for 3 hours and in B: 30°C overnight. FLAG-protein-SUMO1 FLAG-protein Samples were separated on 10% SDS-PAGE and transferred to PVDP membrane by semidry blotting. Membrane was blocked with 5% skim milk in PBST (0.01% Tween 20) for 1h in RT with agitation. Then blot was incubated with primary antibody anti FLAG at dilution of 1:5000 in 5% skim milk/PBST for 1h in RT with agitation. After washes (3 times for 5 min. in PBST) blot was incubated with secondary antibody (anti rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) at dilution 1:25 000 for 1h in RT also with 5% skim milk. The blot was washed 3 times for 5 min. with PBST and developed for 2 min with ECL according to the manufacturer's instructions. Exposure time was 1.5 min.

Dr Agata Cieśla, Ludwików's lab, Laboratory of Biotechnology UAM, Poland

Additional information

Background

Background Rabbit anti-DYKDDDDK is a primary antibody which binds to DYKDDDDK (Sigma FLAG®).

Product citations

immunogen:	KLH-conjugated synthetic peptide: DYKDDDDK (Sigma FLAG®)
Reconstitution:	For reconstitution add 50 µl, of sterile water.
Host:	Rabbit
Clonality:	Polyclonal
Purity:	Immunogen affinity purified serum, in PBS pH 7.4.
Format:	Lyophilized
Quantity:	50 µg
storage:	Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	Western blot (WB)
recommended dilution:	1 : 1000 - 1: 5000 (WB)
calculated \| apparent molecular mass [kDa]:	Depends upon fusion protein partner
Confirmed reactivity:	DYKDDDDK epitope tag (Sigma FLAG®), fused to proteins in plant cells
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	Recombinant FLAG-protein was expressed in bacteria and purified with affinity chromatography. 1.5 µg of FLAG protein was used per in vitro SUMOylation assay in two different conditions: line A. in 30°C for 3 hours and in B: 30°C overnight. FLAG-protein-SUMO1 FLAG-protein Samples were separated on 10% SDS-PAGE and transferred to PVDP membrane by semidry blotting. Membrane was blocked with 5% skim milk in PBST (0.01% Tween 20) for 1h in RT with agitation. Then blot was incubated with primary antibody anti FLAG at dilution of 1:5000 in 5% skim milk/PBST for 1h in RT with agitation. After washes (3 times for 5 min. in PBST) blot was incubated with secondary antibody (anti rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) at dilution 1:25 000 for 1h in RT also with 5% skim milk. The blot was washed 3 times for 5 min. with PBST and developed for 2 min with ECL according to the manufacturer's instructions. Exposure time was 1.5 min. Dr Agata Cieśla, Ludwików's lab, Laboratory of Biotechnology UAM, Poland
background:	Rabbit anti-DYKDDDDK is a primary antibody which binds to DYKDDDDK (Sigma FLAG®).