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Immunofluorescent analysis of GST was performed on HEK-293 cells transiently overexpressing GST - GAD65. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ GST recombinant rabbit monoclonal antibody (Product # 700775) at a dilution of 1:400 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e is untransfected HEK-293 cells. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Tag|
|Published species reactivity||Plant|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein corresponding to amino acids 1-220.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
GST (Glutathione S-transferase) is a 26kDa protein, present in eukaryotes and in prokaryotes, where they catalyze a variety of reactions. Glutathione Stransferase is used to create the GST gene fusion system. This GST-fusion protein can then be purified from cells via its high affinity for glutathione. Fusion Proteins offer an important biological assay for direct protein-to-protein interactions. The GST gene fusion system has been introduced into numerous expression vectors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Arabidopsis CaM binding protein CBP60g contributes to MAMP-induced SA accumulation and is involved in disease resistance against Pseudomonas syringae.
||Wang L,Tsuda K,Sato M,Cohen JD,Katagiri F,Glazebrook J||PLoS pathogens (5:null)||2009|
Glutathione-S-Transferase, GST protein, GST Tag