Product Pathways - Chromatin Regulation / Epigenetics
Sirtuin Antibody Sampler Kit #9787
|9787S||1 Kit (7 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-SirT1 (Ser47) Antibody #2314||40 µl||W, IP, IF-IC, F||H||120||Rabbit|
|SirT1 (D1D7) Rabbit mAb #9475||40 µl||W, IF-IC||H, M, R, Mk||C, B, Pg, Hr||120||Rabbit IgG|
|SirT2 (D4O5O) Rabbit mAb #12650||40 µl||W||H, M, R, Mk||39, 43||Rabbit IgG|
|SirT3 (D22A3) Rabbit mAb #5490||40 µl||W, IP||H, M, R||28||Rabbit IgG|
|SirT5 (D8C3) Rabbit mAb #8782||40 µl||W||H, M, R||30||Rabbit IgG|
|SirT6 (D8D12) Rabbit mAb #12486||40 µl||W, IP, IF-IC||H, M, R, Mk||42, 36||Rabbit IgG|
|SirT7 (D3K5A) Rabbit mAb #5360||40 µl||W, IP||H, M, R, Mk||45||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
Western blot analysis of extracts from various cell lines using SirT6 (D8D12) Rabbit mAb #12486.
Western blot analysis of extracts from various cell lines using SirT2 (D4O5O) Rabbit mAb #12650.
Western blot analysis of extracts from 293 cells, untreated (-) or λ phosphatase-treated (+), using Phospho-SirT1 (Ser47) Antibody #2314 (upper) or SirT1 Antibody #2310 (lower).
Western blot analysis of extracts from various cell lines using SirT7 (D3K5A) Rabbit mAb #5360.
Western blot analysis of mitochondrial extracts from SirT3 wild-type (WT) and knockout (KO) mouse liver, and of whole cell extracts from rat liver and human kidney, using SirT3 (D22A3) Rabbit mAb #5490 (upper) and AIF (D39D2) XP® Rabbit mAb #5318 (lower).
Western blot analysis of extracts from 293 cells expressing either nontargeting shRNA (293 shNT) or shRNA targeting SirT5 (293 shSirT5), and HCT 116, Raw 264.7, and H-4-II-E cells using SirT5 (D8C3) Rabbit mAb #8782 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). 293 shNT and 293 shSirT5 cells were kindly provided by David Lombard, University of Michigan.
The Sirtuin Antibody Sampler Kit provides an economical means of evaluating total levels of sirtuin proteins. The kit includes enough antibody to perform at least four western blot experiments with each primary antibody.
Specificity / Sensitivity
Each antibody in the Sirtuin Antibody Sampler Kit recognizes endogenous levels of its specific target. Activation state antibodies detect their intended targets only when phosphorylated at the indicated site.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Phe297 of human SirT1 protein, Pro205 of human SirT2, or Val130 of mouse SirT3 protein, and with recombinant proteins specific to full-length human SirT5 protein, full-length mouse SirT6, or the amino terminus of human SirT7 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser47 of human SirT1. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.
The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as Class III histone deacetylases. The best characterized of these genes is Saccharomyces cerevisiae Sir2, which is involved in silencing of mating type loci, telomere maintenance, DNA damage response and cell aging (1). SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. SirT2, one of several mammalian Sir2 homologs, deacetylates α-tubulin on Lys40 and histone H4 on Lys16, and is implicated in cytoskeletal regulation and progression through mitosis (2,3). SirT2 protein is mainly cytoplasmic and is associated with microtubules and the HDAC6 tubulin deacetylase (2).
SirT3 exists in human cells in two forms, including a full-length, nuclear (44 kDa) protein and a processed (28 kDa) protein found exclusively in the mitochondria (4-6). Full-length SirT3 protein is processed in the mitochondrial matrix by mitochondrial matrix processing peptidase (MMP) (5). Both full-length and processed SirT3 are active enzymes that deacetylate histone H3 at Lys9 and histone H4 at Lys16 in vitro (4). SirT3 also deacetylates Lys642 of acetyl-CoA synthetase 2 (AceCS2) and activates AceCS2 activity in the mitochondria (7).
SirT5 is localized to the mitochondria and has been implicated in the regulation of cell metabolism (8,9). Nuclear SirT6 is a chromatin-associated protein that promotes normal maintenance of genome integrity as mediated by the base excision repair (BER) pathway (10-12). Mammalian SirT7 is localized to the nucleolus and is prominently expressed in hematopoietic cells, especially myeloid progenitor cells (13). SirT7 is recruited to chromatin by sequence-specific DNA binding transcription factors such as Elk-4, where it facilitates transcriptional repression through deacetylation of histone H3 at Lys18 (14).
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XP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.