GST-tag (mouse, monoclonal)

AS21 4677 | Clonality: Monoclonal | Host: Mouse | Reactivity: GST-tagged proteins

Product Information

Immunogen GST (glutathione S-transferase) recombinant protein

Host Mouse

Clonality Monoclonal

Subclass/isotype IgG1a

Purity Affinity purified in PBS pH 7.4. Contains 0.02 % sodium azide. Contains 50 % glycerol.

Format Liquid

Quantity 50 µg

Storage Store at -20°C for up to 3 years, 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: 500 - 1: 5000 (WB)

Expected | apparent MW Depends upon MW of a fusion protein

Reactivity

Application examples

Application examples Western blot detection using anti-GST monoclonal antibodies

Proteins were isolated from E. coli Rosetta cells transformed with pGEX6P-1 vector. Lane 1 contains protein fraction from bacterial cells before induction with IPTG (-). Lane 2 contains protein fraction from bacterial cells after 2 h induction with IPTG (+). 15uL of sample were loaded and separated on an 15 % SDS-PAGE and transferred for 70 min at 55V using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1x PBS + 0.1 %Tween 20 (T) + 5% milk for 1 h at room temperature (RT) with agitation. Blots were incubated with the primary antibody α-GST (AS21 4677) at indicated dilutions overnight at 4°C with agitation in 1x PBS-T + 5% milk. The primary antibody solutions were decanted, and the blots were washed 4 times (6-8 minutes each) in 1x PBS-T at RT with agitation. Blots were incubated with secondary antibody Goat anti-Mouse (Product no: AS09 627-trial) diluted to 1 : 20,000 in 1x PBS-T + 5% milk for 2 h at RT with agitation. The blots were washed as above and developed with ECL or chemiluminescent detection reagents AgriseraECL SuperBright (AS16 ECL-S-10) for 4 min and imaged using ChemiDoc (BioRad) for 5 seconds. PonceauS served as loading control.

Courtesy of Kelly Mason and Antje Heese; Dept. Biochemistry, IPG; University of Missouri, Columbia (MO), USA

Additional information

This antibody is recognizing native and denatured forms of GST

Background

Background GST-tag (glutathione S-transferase) is a tag added to a protein of interest as a fusion protein to unable purification and detection. The tag is 220 amino acid long, ca. 26 kDa which makes it quite large compare to Myc or FLAG tags.

Product citations

additional information (application):	This antibody is recognizing native and denatured forms of GST
calculated \| apparent molecular mass [kDa]:	Depends upon MW of a fusion protein
Clonality:	Monoclonal
Format:	Liquid
Host:	Mouse
immunogen:	GST (glutathione S-transferase) recombinant protein
Purity:	Affinity purified in PBS pH 7.4. Contains 0.02 % sodium azide. Contains 50 % glycerol.
Quantity:	50 µg
recommended dilution:	1: 500 - 1: 5000 (WB)
storage:	Store at -20°C for up to 3 years, 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.
Sub class:	IgG1a
tested applications:	Western blot (WB)
Picture (footer):	Proteins were isolated from E. coli Rosetta cells transformed with pGEX6P-1 vector. Lane 1 contains protein fraction from bacterial cells before induction with IPTG (-). Lane 2 contains protein fraction from bacterial cells after 2 h induction with IPTG (+). 15uL of sample were loaded and separated on an 15 % SDS-PAGE and transferred for 70 min at 55V using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1x PBS + 0.1 %Tween 20 (T) + 5% milk for 1 h at room temperature (RT) with agitation. Blots were incubated with the primary antibody α-GST (AS21 4677) at indicated dilutions overnight at 4°C with agitation in 1x PBS-T + 5% milk. The primary antibody solutions were decanted, and the blots were washed 4 times (6-8 minutes each) in 1x PBS-T at RT with agitation. Blots were incubated with secondary antibody Goat anti-Mouse (Product no: AS09 627-trial) diluted to 1 : 20,000 in 1x PBS-T + 5% milk for 2 h at RT with agitation. The blots were washed as above and developed with ECL or chemiluminescent detection reagents AgriseraECL SuperBright (AS16 ECL-S-10) for 4 min and imaged using ChemiDoc (BioRad) for 5 seconds. PonceauS served as loading control. Courtesy of Kelly Mason and Antje Heese; Dept. Biochemistry, IPG; University of Missouri, Columbia (MO), USA
background:	GST-tag (glutathione S-transferase) is a tag added to a protein of interest as a fusion protein to unable purification and detection. The tag is 220 amino acid long, ca. 26 kDa which makes it quite large compare to Myc or FLAG tags.