14-3-3 GRF | General regulatory element

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AS12 2119 | clonality: polyclonal | host: rabbit | reactivity: A. thaliana, L. longiflorum. M. crystallinum, algae N. oloabundans


14 st
Item No:
AS12 2119

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product information
Background GRF (General regulatory factor 2) or 14.3.3’s are 30KDa proteins involved in protein interactions with target proteins containing phosphorylated target sites. Functions of 14.3.3’s include acting as adaptors or scaffolds, stimulating protein-protein interaction, altering target protein activity, causing conformational changes of target proteins, regulating subcellular localisation and also facilitating transport (for example nuclear import/export and transport in the endomembrane system). A variety of apparently unrelated biological activities, including a role in development and signal transduction - brassinosteroid mediated signaling pathway.
Immunogen conserved surface-exposed peptide conjugated to KLH.This sequence is repeated in all 12 “canonical” members of the Arabidopsis thaliana GRF/14-3-3 family. GRF1 GF14 chi UniProt: F4JJ94, TAIR: AT4G09000,GRF2 GF14 omega, UniProt:Q01525, TAIR:AT1G78300, GRF3 GF14 psi UniProt: F4KBI7 , TAIR:AT5G38480,GRF4 GF14 phi UniProt:F4HWQ5, TAIR: AT1G35160,GRF5 GF14 upsilon UniProt: P42645, TAIR:AT5G16050,GRF6 GF14 lambda UniProt: P48349#P48349-2, TAIR:AT5G10450,GRF7 GF14 nu UniProt:Q96300, TAIR:AT3G02520,GRF8 GF14 kappa UniProt:F4KHY7, TAIR:AT5G65430,GRF9 GF14 mu UniProt: F4IP55, TAIR:AT2G42590, GRF10 GF14 epsilon UniProt: F4I1C1, TAIR:AT1G22300, GRF11 GF14 omicron UniProt: Q9S9Z8, TAIR: AT1G34760, GRF12 GF14 iota UniProt:Q9C5W6 TAIR: AT1G26480.
Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution please add 50 ĩl of sterile water

Store lyophilized/reconstituted at -20°C; 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)
Related products

AS08 317 | 14.3.3C | 14.3.3-like protein C, rabbit antibody specific to Hordeum vulgare

matching secondary antibody for chemiluminescence detection

Plant protein extraction buffer

Secondary antibodies

Additional information

This antibody is recognizing recombiant GRF of Lilium longiflorum Lil1433_0 accession: AF191746, Lil1433_2, accession: EF397608 and recombinant GRF1,2,3,5 and 6 of Arabidopsis thaliana: GRF1 14-3-3 chi  (At4g09000.1),
GRF2 14-3-3 omega  (At1g78300.1), GRF3 14-3-3 psi (At5g38480.1),
GRF5 14-3-3 upsilon (At5g16050.1), GRF6 14-3-3 lambda (At5g10450.2),
GRF8 14-3-3 kappa (At5g65430.1), GRF11 14-3-3 omicron (At1g34760.1).
There is also very weak reaction to Physcomitrella patens Pp14-3-3 Pp1s 73_133V6 (closest homolog to AtGRF6) and Chlamydomonas reinhardtii Cr 14-3-3 Cre 12.g559250 (closest homolog to AtGRF6).

application information
Recommended dilution 1: 2000 (WB)
Expected | apparent MW

28-20 kDa (depending upon an isoform)

Confirmed reactivity

Arabidopsis thaliana, Lilium longiflorum, Mesembryanthemum crystallinum., algae Neochloris oloabundans

Predicted reactivity Triticum aestivum, Zea mays, Oryza sativa, Pisum sativum, Nicotiana tabacum, Solanum tuberosum, Gossypium hirsutum, Brassica sp., Physcomitrella patens, Chlamydomonas reinhardtii
Not reactive in

This antibody does not bind to 14-3-3-like protein D of Nicotiana tabacum.

Additional information

to be added when available

Selected references

to be added when available, antibody released in September 2013.

application example

 western blot using anti-plant GRF antibodywestern blot detection using anti-plant GRF antibodies 

Left panel: 150 ng of recombinant Arabidopsis thaliana, Physcomitrella patens and Chlamydomonas reinhardtii GRFs (see more details below)  were separated on 12 % SDS-PAGE and blotted 1h to PVDF by semi dry blotting (0.8 mA/cm2 for 60 min). Blots were blocked with 2% skimmed milk powder dissolved in TBST (0.1 % Tween 20) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in blocking solution and incubated 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-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in  for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Prime western blotting detection reagent according to the manufacturer's instructions (GE Healthcare, #RPN2232). Exposure time was 10 minutes.

Right panel: 50 µg of a total cell extract of Arabidopsis thaliana wilde type. Conditions as above, exposure time 30 seconds.

GRF protein designations: At 14-3-3 lambda (GRF6), At 14-3-3 chi (GRF1), At14-3-3 psi (GRF3), At14-3-3 omega (GRF2), At14-3-3 upsilon (GRF5), At14-3-3 omicron (GRF11), At14-3-3 kappa (GRF8), Pp 14-3-3 Pp1s 73_133V6 (closest homolog to At GRF6), Cr 14-3-3 Cre 12.g559250 (closest homolog to AtGRF6).  Note that all recombinant GRFs were detected by this antibody, At14-3-3 kappa (GRF8) to a lesser extent.

Courtesy of Dr. Bernhard Wurzinger, University of Vienna, Austria

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