Chromatin Immunoprecipitation (ChIP)
(ChIP-Seq of AbFlex Histone H3.3 antibody (rAb) ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from brain tumor cell cells and 4 ug of AbFlex Histone H3.3 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15.1 million sequence tags were mapped to identify Histone H3.3 binding sites. The image shows binding across a region of chromosome 1.)
Mouse anti-Human Histone H3.3 Antibody | anti-H3.3 antibody
AbFlex Histone H3.3 antibody (rAb)
Reactivity
Human
Applications
Chromatin Immunoprecipitation, Immunoprecipitation, Western Blot
Purity
Protein A Chromatography
Synonyms
Histone H3.3; AbFlex Histone H3.3 antibody (rAb); anti-H3.3 antibody
Host
Mouse
Reactivity
Human
Isotype
IgG2a
Purity/Purification
Protein A Chromatography
Form/Format
Purified IgG in 140mM Hepes, pH7.5, 70mM NaCl, 32mM NaOAc, 0.035% sodium azide, 30% glycerol. Sodium azide is highly toxic.
Concentration
1ug/ul (varies by lot)
Applicable Applications for anti-H3.3 antibody
Chromatin Immunoprecipitation-Seq (ChIP-Seq), Western Blot (WB)
Application Notes
Applications Validated:
ChIP-Seq: 4ug per ChIP
WB: 2ug/ml
Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western Blot.
ChIP-Seq: 4ug per ChIP
WB: 2ug/ml
Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western Blot.
Antibody Type
Recombinant Antibody
Immunogen
This antibody was raised against a peptide corresponding to amino acids 21-39 of human Histone H3.3.
Preparation and Storage
Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20 degree C for up to 2 years. Keep all reagents on ice when not in storage.1ug/ul
Chromatin Immunoprecipitation (ChIP)
(ChIP-Seq of AbFlex Histone H3.3 antibody (rAb) ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from brain tumor cell cells and 4 ug of AbFlex Histone H3.3 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15.1 million sequence tags were mapped to identify Histone H3.3 binding sites. The image shows binding across a region of chromosome 1.)
Chromatin Immunoprecipitation (ChIP)
(ChIP-Seq of AbFlex Histone H3.3 antibody (rAb) ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from brain tumor cell cells and 4 ug of AbFlex Histone H3.3 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15.1 million sequence tags were mapped to identify Histone H3.3 binding sites. The image shows binding across a region of chromosome 1.)
Western Blot (WB)
(AbFlex Histone H3.3 antibody (rAb) tested by Western blot. 20 ug of HeLa cell nuclear extract was run on an SDS-PAGE gel and probed with 2 ug/ml AbFlex Histone H3.3 antibody. MW: 17 kDa)
Western Blot (WB)
(AbFlex Histone H3.3 antibody (rAb) tested by Western blot. 20 ug of HeLa cell nuclear extract was run on an SDS-PAGE gel and probed with 2 ug/ml AbFlex Histone H3.3 antibody. MW: 17 kDa)
Western Blot (WB)
(AbFlex Histone H3.3 antibody (rAb) tested by Western Blot Lane 1: 250 ng of Recombinant Histone H3.1; Lane 2: 250 ng of Recombinant Histone H3.3. Proteins were run on SDS-PAGE, transferred to nitrocellulose and stained with Ponceau S (lower panel) or probed with AbFlex Histone H3.3 antibody (upper panel). The AbFlex antibody only detects, and is specific for, Histone H3.3 as seen by a band at 17 kDa. For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.)
Western Blot (WB)
(AbFlex Histone H3.3 antibody (rAb) tested by Western Blot Lane 1: 250 ng of Recombinant Histone H3.1; Lane 2: 250 ng of Recombinant Histone H3.3. Proteins were run on SDS-PAGE, transferred to nitrocellulose and stained with Ponceau S (lower panel) or probed with AbFlex Histone H3.3 antibody (upper panel). The AbFlex antibody only detects, and is specific for, Histone H3.3 as seen by a band at 17 kDa. For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.)
Related Product Information for anti-H3.3 antibody
Short Description: AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. Our AbFlex Histone H3.3 antibody (rAb) was raised in a Mouse host. It has been validated for use in ChIP-Seq and Western blot; it has been shown to react with Human samples.
Background: AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex Histone H3.3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. Histone H3.1 and Histone H3.3 are the two main Histone H3 variants found in plants and animals. They are known to be important for gene regulation. Histone H3.1 and H3.3 have been shown to demonstrate unique genomic localization patterns thought to be associated with their specific functions in regulation of gene activity. Specifically, Histone H3.1 localization is found to coincide with genomic regions containing chromatin repressive marks (H3K9me3, H3K27me3 and DNA methylation), whereas Histone H3.3 primarily colocalizes with marks associated with gene activation (H3K4me3, H2BK120ub1, and RNA pol II occupancy). Deposition of the Histone H3.1 variant into the nucleosome correlates with the canonical DNA synthesis-dependent deposition pathway, whereas Histone H3.3 primarily serves as the replacement Histone H3 variant outside of S-phase, such as during gene transcription. Aberrant localization of these variants is also known to correlate with certain cancers.
Background: AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex Histone H3.3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. Histone H3.1 and Histone H3.3 are the two main Histone H3 variants found in plants and animals. They are known to be important for gene regulation. Histone H3.1 and H3.3 have been shown to demonstrate unique genomic localization patterns thought to be associated with their specific functions in regulation of gene activity. Specifically, Histone H3.1 localization is found to coincide with genomic regions containing chromatin repressive marks (H3K9me3, H3K27me3 and DNA methylation), whereas Histone H3.3 primarily colocalizes with marks associated with gene activation (H3K4me3, H2BK120ub1, and RNA pol II occupancy). Deposition of the Histone H3.1 variant into the nucleosome correlates with the canonical DNA synthesis-dependent deposition pathway, whereas Histone H3.3 primarily serves as the replacement Histone H3 variant outside of S-phase, such as during gene transcription. Aberrant localization of these variants is also known to correlate with certain cancers.
Product Categories/Family for anti-H3.3 antibody
NCBI and Uniprot Product Information
Molecular Weight
17kDa
Protein Family
Similar Products
Product Notes
The H3.3 (Catalog #AAA389460) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The AbFlex Histone H3.3 antibody (rAb) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Histone H3.3 can be used in a range of immunoassay formats including, but not limited to, Chromatin Immunoprecipitation-Seq (ChIP-Seq), Western Blot (WB). Applications Validated: ChIP-Seq: 4ug per ChIP WB: 2ug/ml Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western Blot. Researchers should empirically determine the suitability of the H3.3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Histone H3.3, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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