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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)

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Product Information

Immunogen KLH-conjugated peptide derived from synthetic peptide, UniProt: A0A1S4NYF2 from common lancelet Branchiostoma lanceolatum
Host Rabbit
Clonality Polyclonal
Purity Antigen 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 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 - 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

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

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.

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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.

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