One-Step^TM IP-Western Kit

Immunoprecipitation (IP) is a common technique used for identifying protein-protein interactions. IP is usually performed using a primary antibody together with a second antibody resin or Protein A or G resin. After IP, a Western blot is used to detect any possible proteins associated with the target protein.

One problem with IP Western blotting, however, is that the image is always contaminated with heavy chains, light chains or Proteins A or G from the resins. This makes the desired proteins harder to identify.

An innovative product from GenScript, the One-Step^TMIP-Western Kit, completely removes this obstacle. With this kit, the user can apply Protein A, G, A/G or any secondary antibody resin with NO contamination whatsoever from heavy chains, light chains or Proteins A or G.

• GenScript has developed a blocking solution (Pretreatment Solution) that selectively binds and blocks both heavy and light chains of primary antibodies and proteins A and G used for IP, preventing cross-reactivity between:

1) The secondary antibody of subsequent western blot and the primary antibody used for IP, and
2) Between primary or secondary antibody of subsequent western blot and the proteins A and/or G used for IP.

• The blocking solution is formulized to bind and block the blot with greatly improved efficiency.This shortens blocking time down to only five minutes.
• The One-Step IP-Western technology integrates the secondary antibody-binding step, needs no secondary antibody.
• The One-Step IP-Western technology simplifies and shortens entire IP-western blot procedure down to only one hour.

One-Step^TM IP-Western Kit contains all the necessary reagents, buffers, nitrocellulose membrane and HRP substrate for performing an IP-Western blot. After transferring the proteins from gel to membrane, the user incubate the membrane in Pretreat Solution for five minutes, follwed by incubation in WB solution (prepared from primary antibody, IP-WB 1, IP-WB 2 and IP-WB 3) for 40 minutes. Then the membrane is washed three times for five minutes each. The blot can be developed using the HRP substrate included in the kit. The One-Step^TM IP-Western blot result is compared with the conventional western result in Figure 1.

One-Step^TM Complete IP-Western Kits including L00231 (for rabbit primary antibody), L00232(for mouse primary antibody) and L00233 (for goat primary antibody) combines high sensitivity with low background. It detects nanograms of the desired antigen while leaving quantities of antibody as high as 2 μg per lane undetected. The A&G blocker included in the kit takes five minutes to use and gives clean IP-Western results even with serious Protein A, G or A/G carryover contamination.For kit L00231 (rabbit primary antibody), Protein G Blocker is simply added to the IP-WB 3 solution. The protein A carryover does not affect the Western results.

Applications

One-Step^TM IP-Western Kit enables the fast Western-blot for detecting proteins from immunoprecipitation using:

• Protein A, G or A/G Resin.
• Any secondary antibody Resin.

	Cat. No.	Name	Size	Price	Quantity
	L00231	One-Step IP-Western Kit (Rabbit)	1 Kit (10 Assays)	$375.00
	L00232	One-Step IP-Western Kit (Mouse)	1 Kit (10 Assays)	$375.00
	L00233	One-Step IP-Western Kit (Goat)	1 kit (10 Assays)	$375.00