GFP | Green fluorescent protein (VENUS)
AS18 4227 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Venus
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Product Information
Immunogen
Full length, recombinat VENUS protein, expressed in E.coli, UniProt: P42212
Host
Rabbit
Clonality
Polyclonal
Purity
Affinity purified serum, in PBS pH 7.4
Format
Lyophilized
Liquid
Liquid
Quantity
50 µg
Reconstitution
For reconstitution add 50 µl, of sterile water.
Storage
Lyophilized antibody can be stored at -20°C for up to 3 years.
Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications
Western blot (WB)
Recommended dilution
1 : 1000 (WB)
Expected | apparent MW
26 kDa
Reactivity
Confirmed reactivity
cell lysate overexoressing Venus protein fusion
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
Application example
200 ng mVenus YFP protein (1); 100 ng mVenus YFP protein (2) ; 49 ng mVenus YFP protein (3) ; 24 ng mVenus YFP protein (4) ; 12 ng mVenus YFP protein (5)
MW markers: BenchMark™ Pre-stained Protein Ladder (10748010)
>0.200 – 0.012 µg of total protein from a pure stock of mVenus YFP protein in 1x SIGMAFAST EDTA-free Protease Inhibitor (S8830-2TAB) and denatured with 1x reducing Laemmli SDS buffer at 90°C for 10 min, and insoluble material pelleted at 20,000 xg for 15 min. Samples were run to separation on a 4-12% SDS-PAGE gel and blotted 7 mins to PVDF using ThermoFisher iBlot high voltage Protocol 3. Blot was blocked with 7.5 % milk in TBS-T for 20mins/RT with agitation. Blot was incubated in the primary antibody (i.e. Rabbit anti-Venus IgG) at a dilution of 1:10 000 with 7.5 % milk in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly three times, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was blocked with 7.5 % milk in TBS-T for 20mins/RT. Blot was then incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in TBS-T for 2.5h/RT with agitation. The blot was washed as above, then as above with TBS, and developed for 5 min with chemiluminescent detection reagent, according to manufature's instruction.
Exposure time was approximately 3 seconds.
200 ng mVenus YFP protein (1); 100 ng mVenus YFP protein (2) ; 49 ng mVenus YFP protein (3) ; 24 ng mVenus YFP protein (4) ; 12 ng mVenus YFP protein (5)
MW markers: BenchMark™ Pre-stained Protein Ladder (10748010)
>0.200 – 0.012 µg of total protein from a pure stock of mVenus YFP protein in 1x SIGMAFAST EDTA-free Protease Inhibitor (S8830-2TAB) and denatured with 1x reducing Laemmli SDS buffer at 90°C for 10 min, and insoluble material pelleted at 20,000 xg for 15 min. Samples were run to separation on a 4-12% SDS-PAGE gel and blotted 7 mins to PVDF using ThermoFisher iBlot high voltage Protocol 3. Blot was blocked with 7.5 % milk in TBS-T for 20mins/RT with agitation. Blot was incubated in the primary antibody (i.e. Rabbit anti-Venus IgG) at a dilution of 1:10 000 with 7.5 % milk in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly three times, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was blocked with 7.5 % milk in TBS-T for 20mins/RT. Blot was then incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in TBS-T for 2.5h/RT with agitation. The blot was washed as above, then as above with TBS, and developed for 5 min with chemiluminescent detection reagent, according to manufature's instruction.
Exposure time was approximately 3 seconds.
Courtesy of Dr. Dr Nanakow Baiden at Rothamsted Research, UK
Additional information
Background
Background
Green fluorescent protein (Venus) from Aequorea victoria (Jellyfish), can be mutated to emit at different wavelengths such as blue for BFP (when Tyr-66 is replaced by His), cyan for CFP (when Tyr-66 is replaced by Trp), and yellow for YFP (when THR-203 is replaced by Tyr). It is an energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer.
Product citations
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