ChIP Grade and ChIP Validated Antibodies

Antibodies are essential tools in many epigenetics and non-coding RNA experiments. This database contains only antibodies that have been qualified and/or validated to work in ChIP, RIP, and MeDIP applications. Suppliers vary in how they define “qualified,” “grade,” and “validated,” so we’ve outlined the definitions for these within this database in the Validation section.

Chromatin

Despite recent high-throughput and technology advances, antibodies remain the backbone of widely used techniques, such as chromatin immunoprecipitation (ChIP) for studying chromatin structure and function. The antibodies in this section are targeted against the epitopes of chromatin modifying proteins, histone deacetylase or demthylase enzymes, as well as histones and their various modifications. They can be used to explore the role of chromatin in epigenetics and gene regulation.

DNA Methylation

Antibodies play a key role in widely used techniques for DNA methylation quantification and location analysis. The antibodies in this section are targeted against methylated DNA including several variants like 5-hydroxymethylcytosine (5-hmC), 5-carboxylcytosine (5-caC) and 5-formylcytosine (5-fC), as well as several proteins involved in the DNA methylation process, in order to study the role of DNA methylation in epigenetics and gene regulation.

Non-Coding RNA

Antibodies that bind key RNAi proteins and complexes like Dicer, the RISC complex, and Argonaute family have been extremely useful in gaining a better understanding of RNA regulation. The RNAi related antibodies in this section are targeted against miRNA related and RNA binding proteins. They can be used in detection or pulldown experiments like RNA Binding Protein Immunoprecipitation (RIP) as a way to elucidate or characterize non-coding RNA mechanisms.

ChIP Grade Antibodies

ChIP Grade antibodies have typically been validated by an antibody supplier or another external group to work well in ChIP experiments. Although suppliers almost always validate their antibodies, they can't do so with every application where the antibody may be used. As a result, most antibodies will only have a subset of their offering validated as ChIP Grade. This doesn't mean that antibodies that aren't ChIP Grade will not work. It only means that have not yet been validated in that application.

ChIP Validated Antibodies

ChIP Validated Antibodies are usually validated by an antibody supplier to work well in ChIP applications. Unlike ChIP Grade antibodies, ChIP Validated antibodies undergo this functional QC for every lot of antibody produced. This higher level of QC results in fewer antibodies available under this category.

Chromatin Modifying Antibodies

Antibodies to chromatin related proteins involved in the modification of chromatin structure and function. These antibodies are often used in chromatin immunoprecipitation (ChIP) experiments.

DNA Methylation and DNA Binding Antibodies

DNA Methylation and DNA Binding Antibodies bind to either methylated DNA motifs or target specific components involved in the DNA methylation pathway. DNA Methylation Antibodies can be used for detection and identification experiments, Immuno-precipitation (IP) experiments, as well as DNA Affinity Reagents for DNA methylation analysis, (i.e. MeDIP) procedures.

Histone Deacetylase Antibodies

Histone deacetylase (HDAC) enzymes influence transcription by selectively deacetylating the ε-amino groups of lysine located near the amino termini of core histone proteins. Chromatin deacetylation correlates with gene silencing. HDACs are also involved in the reversible acetylation of non-histone proteins. Altered HDAC activities are present in many types of cancers. HDAC Antibodies can be used to monitor cellular levels of HDAC enzymes.

Histone Demethylase Antibodies

Antibodies to histone demethylases like the Lysine Specific Demethylase 1 (LSD1) or Jumonji domain-containing (JmjC) enzyme families. Histone demethylation is usually thought of as a permanent modification, but recent studies challenge that notion.

Histone H1 Antibodies

Histone H1 Antibodies bind to Histone H1.Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction.

Histone H2 Antibodies

Histone H2 (including Histone 2A and 2B) Antibodies bind to Histone H2A.Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction.

Histone H3 Antibodies

Histone H3 Antibodies bind to Histone H3. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction.

Histone H4 Antibodies

Histone H4 Antibodies bind to Histone H4. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction.

RNAi Related Antibodies

Antibodies that bind to Dicer, RISC proteins, the Argonaute family and other key players in RNAi processes are key to better understanding this important regulatory pathway.

Monoclonal

Monoclonal antibodies detect only one epitope on a particular antigen. They also consist of only one antibody subtype. Generally, monoclonal antibodies take longer, and require more effort to produce than polyclonal antibodies, however, once hybridomas are made, they can be continually reused to create identical batches; which is helpful for consistent experimental results and standardizing procedures. Monoclonal antibodies usually have less background, and won’t cross-react with other proteins making them suited to use as the primary antibody in assays, detecting antigens in tissue, staining, or affinity purification. Sometimes these attributes may also cause issues; for instance if the antibody is too specific, and won’t work in a broad range of species, or is no longer effective after the loss of an epitope through chemical treatment of the antigen.

Polyclonal

Polyclonal antibodies recognize multiple epitopes on one antigen. They contain a heterogeneous mixture of antibodies with varying affinities. Polyclonal antibodies are relatively quick and inexpensive to produce, but they can be non-specific and encounter lot-to-lot variability. Polyclonal antibodies are suited for certain situations, like amplifying signal from a low expression target protein. Recognition multiple epitopes, makes them useful in immuneprecipitation (IP) or chromatin immunoprecipitation (ChIP) experiments, and they are tolerant of minor changes in antigens, such as polymorphisms, heterogeneity of glycosylation, or slight denaturation. Polyclonal antibodies can also be used to identify proteins similar to the immunogen protein across tissues or species, and are preferred for detection of denatured proteins. The downside to polyclonal antibodies is that there can be high variability between batches, they sometimes give high background signals, and their multiple epitopes make it important to check for cross-reactivity. 

Chicken

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Chicken, so you’ll want to make sure that is appropriate for your application.

Goat

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Goat, so you’ll want to make sure that is appropriate for your application.

Mouse

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Mouse, so you’ll want to make sure that is appropriate for your application.

Rabbit

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Rabbit, so you’ll want to make sure that is appropriate for your application.

Rat

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Rat, so you’ll want to make sure that is appropriate for your application.

Sheep

Antibodies are generated in a wide range of host species. They are usually selected with a host species that matches the species you are studying, but they may also react with a similar target protein from another species if they share enough of the same amino acid sequence. The antibodies in this section were raised in Sheep, so you’ll want to make sure that is appropriate for your application.

Affinity Purified

Affinity purification isolates proteins with similar characteristics. The technique is based on the interactions between the proteins and a specific ligand coupled to a chromatographic matrix and takes advantage of the affinity of the specific immunoglobulin fraction for the antigen it was created to target. The result is the elimination of most of the non-specific immunoglobulin fraction, while enriching the fraction that reacts with the target antigen. The remaining purified material will contain the immunoglobulin of desired specificity.

Ammonium Sulfate Precipitation

Ammonium Sulfate Precipitation

Ascites

Ascites

Culture Supernatant

Culture Supernatant

Diluted Culture Supernatant

Diluted Culture Supernatant

IgG Fraction

IgG fraction

Protein A Purfied

Protein A Purfied

Protein A purified

Protein A purified

Protein G Purified

Protein G purified

Purified

Purified

Serum

Serum

Unpurified

Antibodies are often found in relatively unpurified formats, described as “serum” or “antiserum” (referring to the blood from an immunized host.) The antiserum will contain antibodies/immunoglobulins of all classes as well as other serum proteins. In addition to antibodies that recognize the target antigen, the antiserum also contains antibodies to various non-target antigens that can sometimes react non-specifically in immunological assays. Unpurified antibody preparations vary significantly in specific antibody concentration.

ChIP Grade

ChIP Grade antibodies are those that have either been initially validated by a supplier's R&D team to work well in ChIP applications, or those where researchers have successfully published using them in ChIP experiments. The primary difference between ChIP Grade and ChIP Validated antibodies in this database is that ChIP Validated antibodies are typically validated lot by lot to ensure that each batch of antibody will perform well in ChIP.

ChIP Validated

ChIP Validated antibodies undergo a QC to ensure that they perform well in ChIP applications. This QC for ChIP Validated antibodies is typically performed for every single lot of antibody produced. This higher level of QC results in fewer antibodies available under this category.