Product

Western Blotting

StripPRO™ 1 Min Stripping Buffer
For fast and convenient removal of antibodies

StripPRO™ 1 Min Stripping Buffer effectively removes antibodies from Western blots in one minute. The unconjugated antigens on the stripped membrane are allowed to be reprobed and be detected with chemiluminescent substrates. StripPRO™ 1 Min Stripping Buffer is an ideal product for breaking antigen-antibody interaction, saving time and saving conserving samples.
产品信息

Highlights:

 

  • Extreme short working time: only one minute stripping to remove primary and secondary antibodies
  • Harmless formula: free of reducing agents, toxic compounds and odors

 

Order Information:

 

Cat. No. Product Name Description
SP01-500

StripPRO™ 1 Min Stripping Buffer

500 mL Solution X 1 

SP05-100 StripPRO™ 1 Min Stripping Buffer (5X)

100 mL Solution X 1

 

 

Product Detail:

 

 

 

Figure 1. StripPRO™ 1 Min Stripping Buffer removes antibodies from blot in only 1 min

Huh7 cell lysate was probed for ACC (280kD). Blots were then stripped with either StripPRO™ (1min) or brand T (15 mins). The blots were then re-blocked and reprobed for GAPDH (36kD) and detected with LumiFlash™ Ultima Chemiluminescent Substrate.

 

 

 

Figure 2. Stripping and reprobing blot for different molecular weight targets with StripPRO™

 

1/2 fold serial dilution of Hela cell lysate (start from 20ug/well) were separated by SDS-PAGE. The proteins were transferred to 0.45um PVDF membranes and the blocked with BlockPRO™ 1 Min Protein-Free Blocking buffer (BM01) and analyzed by Wetsern blot using LumiFlash™ Ultima Chemiluminescent substrate, HRP system (LF08) and the Chemlux SPX-600V (2x2 binning).  The first target (1st detection) was detected at 1:1,000 by probing with Anti-ACC monoclonal antibody (#3676, Cell Signaling Technology). And then the blot was stripped in  1 Min Stripping Buffer (SP01) for 1 minute at room temperature, washed in 1X TBS Tween-20, incubated with substrate and imaged to check for stripping efficiency (1st strip test) The second target (2nd detection) was detected at 1:1000 by reblocking and probing with Anti-p38 monoclonal antibody (ab31828, Abcam).  The blot was stripped again (2nd strip test) and then the third target (3rd detection) was detected at 1:10,000 by reblocking and probing with Anti-GAPDH antibody (ab8245, Abcam). All of the secondary antibodies was proved at 1:10,000 (#115-035-003, 3115-035-144, Jackson ImmunoResearch) 

 

 

 

 

 

Figure 3. Demonstration of phosphorylated protein and original type in one blot by using StripPRO™ 1 Min Stripping Buffer

Hela cell lysate was extracted with RIPA Cell Lysis Buffer (RP05) added with PhosSTOP (Roche). 1/2 fold serial dilution of Hela Cell lysate (start from 20ug/well) were separated by SDS-PAGE. The two membranes were probed for p-ACC and GAPDH respectively and detected with LumiFlash™ Ultima Chemiluminescent substrate, HRP system (LF08). Blots were stripped in StripPRO™ 1 Min Stripping Buffer (SP01) for 1 minute at room temperature. The blots were reblocked and reprobed for ACC and GAPDH respectively.

 

 

 

Figure 4. Stripping efficiency of StripPRO™ 1 Min Stripping Buffer in two common transfer membranes

1/2 fold serial dilution of Hela Cell lysate (start from 20ug/well) were separated by SDS-PAGE. Following protein transferred to nitrocellulose (NC) paper and PVDF, the two membranes were probed for ACC and detected with  LumiFlash™ Ultima Chemiluminescent substrate, HRP system (LF08). Blots were stripped in StripPRO™ 1 Min Stripping Buffer (SP01) for 1 minute at room temperature. The blots were reblocked and reprobed for GAPDH.