Name:
Phone:
E-mail:
Address:






mNeonGreen | Fluorescent Protein

AS21 4525  | Clonality: Polyclonal |  Host: Rabbit | Reactivity: mNeonGreen | Fluorescent Protein


mNeonGreen |  Fluorescent Protein in the group Tag Antibodies / GFP/YFP/RFP/CFP at Agrisera AB (Antibodies for research) (AS21 4525)
mNeonGreen |  Fluorescent Protein



DATA SHEET IN PDF

Qty: 
292
Buy 2 items of this product for 2180.00 €/items
Buy 3 items of this product for 198.00 €/items
How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen KLH-conjugated peptide derived from synthetic peptide, UniProt: A0A1S4NYF2 from common lancelet Branchiostoma lanceolatum
Host Rabbit
Clonality Polyclonal
Purity Immunogen affinity purified serum, in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
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 used fusion partner

Reactivity

Confirmed reactivity mNG tag in plant (Arabidopsis thaliana) and algal vectors (Chlamydomonas reinhardtii)
Predicted reactivity Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Western blot using anti-mNeonGreen antibodies with reactivity to plant and algal fusions
Samples: 
1- Precision Plus Protein Standards (Bio-Rad)
2-Arabidopsis thaliana expressing mNeonGreen-tagged fusion protein
3-Arabidopsis thaliana wildtype total cell extract
4-Chlamydomonas reinhardtii expressing mNeonGreen-tagged fusion protein
5-Chlamydomonas reinhardtii wildtype cell extract

20 µg of total protein extracted from Arabidopsis thaliana leaves or Chlamydomonas reinhardtii cells with 62.5 mM Tris-HCl pH 6.8, 10 % glycerol, 2 % SDS, 50 mM DTT, 1x protease inhibitors (P9599, Sigma) and heated at 65ºC for 5 minutes were separated on 10 % SDS-PAGE and blotted 1 h to PVDF membrane (pore size 0.45 µm) using wet transfer. The blot was blocked with 5 % milk/TBS-T for 1 h/RT with agitation and then incubated in the primary antibody at a dilution 1: 1000 in 5 % milk/TBS-T for 1h/RT with agitation. The antibody solution was decanted, and the blot was briefly rinsed, then washed three times for 10 minutes in TBS-T with agitation. The blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG, HRP conjugated, AS09 602) at a dilution of 1: 25 000 in 5 % milk/TBS-T for 1h/RT with agitation. The blot was washed as above, developed for 2 minutes with AgriseraECLSuperBright substrate and imaged with a Bio-Rad ChemiDoc MP digital imaging system. Exposure time was 2.5 s. 

Courtesy of Hanie Khorshidi, University of Saskatchewan, Canada




Western blot using anti-epitope tag, mNeonGreen antibodies


Samples: 
1 - Nicotiana benthamiana whole leaf extract expressing mNeonGreen-ATG8
2 – Nicotiana benthamiana whole leaf extract expressing YFP-ATG8
3 – Nicotiana benthamiana whole leaf extract expressing mCherry-ATG8

MW markers: (Biorad)Precision Plus Protein Dual Color Standards, 500 µl #1610374

50 µg of total protein extracted freshly from Nicotiana benthamiana leaves with exaction buffer (50mM Tris-HCl pH7.5, 150mM NaCl, 10%(v/v) glycerol, 0.5mM EDTA and 0.5% NP40) denatured with SDS-sample buffer at 95°C for 5 min, proteins were separated on 10 % SDS-PAGE and blotted 0.5h to PVDF membrane using semi-dry transfer. Blots were blocked with 5 % milk in PBS-T for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 3 000 for ON/4°C 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 PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG (H&L), horse radish peroxidase conjugated, AS09 602) diluted to 1: 5 000 in for 1h/RT with agitation. The blot was washed as above, developed for 5 mins with Agrisera ECL Bright and detected with film.

Courtesy of Dr. Jiaqi Sun, McGill University, Canada

Western blot using mNeonGreen antibodies


Samples:

1- PageRuler™ Prestained Protein Ladder (Thermo Fisher)
2- 20 µg Arabidopsis thaliana wildtype microsomal membrane preparation
3- 20 µg Arabidopsis thaliana expressing VHA-a1-GSL-NeonGreen microsomal membrane preparation
4- 20 µg Arabidopsis thaliana expressing VHA-a3-GSL-NeonGreen microsomal membrane preparation

20 µg of microsomal membrane proteins were prepared from Arabidopsis thaliana 4-week -old rosette leaves with a buffer containing 350mM sucrose, 70mM Tris-HCl pH8, 10% glycerol, 3mM Na2EDTA, 0.15% BSA, 1.5% PVP-40, 4mM DTT, 1x Roche completeTM protease Inhibitor and denatured at 95ºC for 5 minutes. Proteins were separated on 10 % SDS-PAGE and blotted for 1 h to PVDF membrane (pore size 0.2 µm) using wet transfer. The blot was blocked with 5% milk/TBS-T (0.05%) for 1 h at RT with agitation and then incubated in the primary antibody at a dilution 1: 1000 in 5% milk/TBS-T (0.05%) for 1h at RT with agitation. The antibody solution was decanted, and the blot was briefly rinsed, then washed three times for 10 minutes in TBS-T (0.05%) with agitation. The blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG, HRP conjugated, AS09 602) at a dilution of 1: 25 000 in 5 % milk/TBS-T (0.05%) for 1hr at RT with agitation. The blot was washed as above, developed for 1 min with AgriseraECLBright substrate and imaged with a cooled CCD camera system (Intas ADVANCED Fluoreszenz u. ECL Imager). Exposure time was 10s.

Courtesy of Upendo Lupanga, Centre for Organismal Studies, University of Heidelberg, Germany

Additional information

Additional information The peptide used to elicit this antibody is conserved in the following expression vectors:

  • dCas9-P2A-DHFR(TSc3)-T2A-mNeonGreen [Cloning vector pBM020]
  • Cas9-P2A-DHFR(TSc3)-T2A-mNeonGreen [Cloning vector pBM006]
  • mNeonGreen4-tDeg [Cloning vector pminiCMV-mNeonGreen4-tDeg]
  • ER-localized mNEONGREEN [Binary vector pKT-NM-erNEON]
  • mNeonGreen-3xFLAG [Cloning vector pLM160-mNeonGreen]
  • mNeonGreen-ty1 [Cloning vector pSAG1-mNeonGreen_TUB1-dTomato]
  • mNeonGreen, partial [Binary vector pRATIO2131]

The peptide used to elicit this antibody is not conserved in GFP protein sequence.
This antibody is detecting protein sequence of mNG tag in plant and algal vectors.

This antibody is also reacting to some degree with YFP and mCherry.

Related products

Background

Background mNeonGreen (Fluorescent Protein) is a new bright monomertic yellow-green fluorescent, with low conservation level to GFP.  It has an excitation maximum at 506 nm and an emission maximum at 517 nm and therefore is compatible with the most GFP filter sets. mNeonGreen is 3x brighter compare to GFP, more stable and does not bleach so fast as GFP, which makes it very suitable for confocal and super resolution microscopy, for fusion proteins with low expression levels. It can be used in bicistronic vectors, which will allow simultaneous expression of two proteins separately, from the same RNA transcript. mNeonGreen has MW of 26.6 kDa.

Product citations

Related products: mNeonGreen | Fluorescent Protein

AS16 ECL-S-N | low pico to mid femtogram and extreme low femtogram detection

This product can b...
From 25 €
AS18 4175  | Clonality: Monoclonal  |  Host: Mouse  |  Reactivity: GFP
320 €
AS09 602 |  Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)
194 €
AS18 4178 | Clonality: Polyclonal | Host: Chicken | Reactivity: mCherry
320 €