Protein-Protein- / Protein-DNA-Interaction

Fast: (co-)immunoprecipitation in just 1,5 hours
Reliable: high specificity due to low background binding
Flexible: high-throughput, automated or low-throughput, manual processing

IP, Co-IP or ChIP of virtually any protein

The µMACS™ and MultiMACS™ Protein A/G Kits are designed for analytical scale immunoprecipitation (IP), Co-IP, or chromatin-IP (ChIP) of proteins and their interaction partners. The non-sedimenting µMACS™ Protein A/G MicroBeads ensure rapid reaction kinetics which allows for the formation of the labeled immune complex in only 30 minutes. Compared to the standard protocol the short protocol and the efficient on-column washing steps help reducing non-specific background binding and potential protein dissociation from the complex.

Fast - Save time on pre-clearing and incubation steps and complete (co)-IP experiments in just 1,5 hours.

Convenient - Cut out multiple centrifuge and buffer removal steps.

Low background - µMACS Protein A/G MicroBeads with low non-specific binding in combination with efficient and gentle on-column washing steps significantly reduce background (see Fig. 2).

Flexible - Easily adaptable from manual low-throughput to automated high-throughput processing by using the MultiMACS^TM M96 Separator.

Chromatin Immunoprecipitation

The advantages of µMACS Protein A/G MicroBeads for (co-)IP have been incorporated into a special protocol for ChIP. In contrast to the standard ChIP protocol this streamlined protocol saves 90% of laboratory time in particular on pre-clearing, incubation and reverse crosslink steps. Customer data indicated an enhanced chromatin immunoprecipitation and much reduced level of non-specific PCR products (see Fig. 4). In addition this special protocol allows for working with just 10⁶ cells as starting material. Please click here to download the protocol. To see an application note reported by David Mulholland and colleagues from the Prostate Research Center, Vancouver, Canada click here.

Hardware

µ Column
μMACS Separator
MultiMACS^TM M96 Separator

Further information

Chromatin immunoprecipitation (ChIP)
[PDF; 94 KB]

Figure 1

Immunoprecipitation using μMACS™ Protein A or Protein G MicroBeads.

Figure 2

Immunoprecipitation of the SV40 large T antigen. SV40 large T antigen was immunoprecipitated from COS cells using μMACS Protein G MicroBeads as shown by silver stained SDS gel of the cell lysate (L), protein marker (M), the immunoprecipitated large T antigen (lane 1, indicated by the arrow), an isotype-matched control antibody (2, rat anti-mouse antibody) and control (3, without antibody).

Figure 3

Co-immunoprecipitation of beta-catenin (BCat). Androgen receptor was immunoprecipitated from dihydrotestosterone (DHT)-stimulated (lane 1, 3) or unstimulated LNCaP cells (2, 4) with an androgen receptor specific antibody using µMACS Protein G MicroBeads (1, 2) or with Protein A/G agarose beads (lanes 3, 4 ). Western blot (WB) using anti-beta-catenin antibody shows BCat co-immunoprecipitated with androgen receptor with higher sensitivity when µMACS Protein G MicroBeads were employed. (Courtesy of D. Mulholland, Vancouver, Canada)

Figure 4

Enhaced chromatin immunoprecipitation of androgen receptor using μMACS Protein G MicroBeads. LNCaP prostate cancer cells were stimulated with dihydrotestosterone (DHT) (lanes 1, 3) or control (lanes 2, 4) for 48 h and ChIP was carried out using an androgen receptor specific antibody and μMACS Protein G MicroBeads (lanes 1, 2) or Protein A/G agarose beads (lanes 3, 4). The experiments carried out with μMACS Protein G MicroBeads showed not only enhanced immunoprecipitation, but also a much reduced level of non-specific PCR product. (Courtesy of d. Mulholland, Vancouver, Canada)