perform ChIP on difficult-to-lyse PBMCs
However, for researchers who want to perform ChIP, PBMCs make it difficult to extract quality chromatin because they are highly resistant to lysis under conditions normally suitable for other cell types. Active Motif leveraged its years of experience working with these difficult-to-lyse cells into developing a kit to finally enable researchers to prepare chromatin and perform ChIP on PBMCs. The ChIP-IT PBMC Kit offers a complete solution for chromatin immunoprecipitation including specialized reagents and protocols to enable extraction of high quality chromatin from PBMCs and integrated components and protocols for ChIP and DNA purification from our highly popular ChIP-IT High Sensitivity® Kit.
To learn more about ChIP-IT® PBMC, click on the Method, Data, or Contents tabs below. To view a manual or other related documents, click on the Documents tab below.
ChIP-IT® PBMC Advantages
- Obtain quality chromatin from PBMCs including T & B lymphocytes, dendritic cells and monocytes
- Sensitive enrichment of DNA from nanogram quantities of chromatin
- Optimized reagents from our ChIP-IT High Sensitivity Kit improve signal and minimize background
- Filtration based washes offer a faster, easier solution with better consistency than magnetic capture
- Highly robust procedure has been validated in both qPCR and ChIP-Seq
Active Motif also offers products compatible with ChIP-IT® PBMC:
- ChIP-IT® qPCR Analysis Kit – simplifies qPCR data analysis and enables normalization across multiple sample types and experiments
- qPCR Primer Sets for a variety of gene targets and model organisms.
- ChIP-Seq Antibodies validated with our ChIP-IT High Sensitivity Kit.
In addition, our Epigenetic Services can perform ChIP, ChIP-Seq and many other genome-wide data generation and analysis services for you.
|ChIP-IT® PBMC||16 rxns||53042||$485||Buy Now|
|ChIP-IT® qPCR Analysis Kit||10 rxns||53029||$95||Buy Now|
|High Sensitivity Chromatin Preparation||16 rxns||53046||$255||Buy Now|
|ChIP-IT® PBMC Kit Manual|
|High Sensitivity Chromatin Preparation Kit Manual|
|Epigenetics Products and Services|
|Tools for Disease Research|
How does ChIP-IT® PBMC work?
Figure 1: Schematic of chromatin immunoprecipitation using ChIP-IT PBMC.
The ChIP-IT® PBMC Kit is specially formulated to perform ChIP from PBMCs, including lymphocytes (T cells, B cells and NK cells), monocytes and dendritic cells. ChIP-IT PBMC offers a complete solution for chromatin immunoprecipitation including specialized reagents and protocols to enable extraction of high quality chromatin from PBMCs and integrated components and protocols for ChIP and DNA purification from our highly popular ChIP-IT High Sensitivity® Kit to reduce background and minimize sample loss for improved sensitivity and consistency.
Figure 1: ChIP-IT PBMC data using chromatin extracted from CD8+ selected T cells showing H3K9ac enrichment at specific loci.
Figure 2: ChIP-IT PBMC utilizes the immunoprecipitation protocol from the ChIP-IT High Sensitivity Kit for improved sensitivity and reproducibility.
ChIP-Seq Validation of ChIP-IT PBMC enriched DNA
The ChIP'd DNA obtained from the ChIP-IT PBMC Kit is of high quality and can be used in downstream applications such as ChIP-Seq or ChIP-chip. Active Motif has validated ChIP-IT PBMC for ChIP-Seq as shown below.
Figure 3: ChIP-Seq data for OGT/O-GlcNAc transferase antibody using chromatin from primary mouse T cells.
Figure 4: ChIP-Seq data for Histone H3K4me2 antibody using chromatin from mouse primary T cells.
Contents & Storage
The ChIP-IT® PBMC Kit contains sufficient reagents to perform 16 chromatin preparation and immunoprecipitation reactions. Reagents for DNA purification are also provided.
The ChIP-IT PBMC Kit is shipped on dry ice and contains reagents with multiple storage temperatures inside. Please confirm receipt of all reagents upon arrival and store items at the appropriate temperatures as indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. Do not re-freeze the Protein G Agarose Beads after you have received this kit. This kit includes the following components:
- RNase A (10 mg/ml); Store at -20°C
- Proteinase K (10 mg/ml); Store at -20°C
- Blocker; Store at -20°C
- 100 mM PMSF; Store at -20°C
- Protease Inhibitor Cocktail; Store at -20°C
- Precipitation Buffer; Store at -20°C
- Carrier; Store at -20°C
- 10X PBS; Store at -20°C
- Protein G Agarose beads; Store at 4°C
- Swelling Buffer; Store at 4°C
- TE, pH 8.0; Store at RT
- Detergent; Store at RT
- Stop Solution; Store at RT
- 5 M NaCl; Store at RT
- ChIP Filtration Columns; Store at RT
- ChIP Buffer; Store at RT
- Wash Buffer AM1; Store at RT
- Elution Buffer AM4; Store at RT
- DNA Purification Binding Buffer; Store at RT
- 3 M Sodium Acetate; Store at RT
- DNA Purification Wash Buffer; Store at RT
- DNA Purification Elution Buffer; Store at RT
- DNA Purification Columns; Store at RT