Revision 1

#9787

Store at -20C

Sirtuin Antibody Sampler Kit

1 Kit

(7 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-SirT1 (Ser47) Antibody231420 µl120 kDaRabbit
SirT1 (D1D7) Rabbit mAb947520 µl120 kDaRabbit IgG
SirT2 (D4O5O) Rabbit mAb1265020 µl39, 43 kDaRabbit IgG
SirT3 (D22A3) Rabbit mAb549020 µl28 kDaRabbit IgG
SirT5 (D8C3) Rabbit mAb878220 µl30 kDaRabbit IgG
SirT6 (D8D12) Rabbit mAb1248620 µl42, 36 kDaRabbit IgG
SirT7 (D3K5A) Rabbit mAb536020 µl45 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

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 two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using SirT6 (D8D12) Rabbit mAb.
undefined Image 1: Sirtuin Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using SirT2 (D4O5O) Rabbit mAb.
undefined Image 2: Sirtuin Antibody Sampler Kit
Western blot analysis of 293 cell lysates treated or untreated with λ phosphatase, using Phospho-SirT1 (Ser47) Antibody #2314 (upper) or SirT1 Antibody #2310 (lower).
undefined Image 3: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using SirT7 (D3K5A) Rabbit mAb.
undefined Image 4: Sirtuin Antibody Sampler Kit
Western blot analysis of mitochondrial extracts from SirT3 wild-type (WT) mouse liver (lane 1) and knockout (KO) mouse liver (lane 2), and of whole cell extracts from rat liver (lane 3) and human kidney (lane 4), using SirT3 (D22A3) Rabbit mAb (upper) and AIF (D39D2) XP® Rabbit mAb #5318 (lower). The absence of signal in the SirT3 knockout mouse liver confirms the specificity of the antibody for SirT3
undefined Image 5: Sirtuin Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 6: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). 293 shNT and 293 shSirT5 cells were kindly provided by David Lombard, University of Michigan.
undefined Image 7: Sirtuin Antibody Sampler Kit
Western blot analysis from wild-type mouse embryo fibroblasts (MEF) (lane 1) or SirT1 knockout mouse embryo fibroblasts (MEF) (lane 2) using SirT1 (D1D7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SirT1 knockout mouse embryo fibroblasts (MEF) confirms specificity of the antibody for SirT1. Wild-type and knockout MEFs were a gift from Wenyi Wei at the Harvard Medical School.
undefined Image 8: Sirtuin Antibody Sampler Kit
Western blot analysis of extracts from HCT 116 cells, expressing either non-targeting shRNA (shNT) or shSirT6, using SirT6 (D8D12) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 9: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from SirT2 wild-type (WT) and knockout (KO) mouse brain using SirT2 (D4O5O) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). SirT2 WT and KO mouse brain extracts were kindly provided by Dr. Gizem Donmez, Tufts University School of Medicine.
undefined Image 10: Sirtuin Antibody Sampler Kit
Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) and phosphatase-treated (right), using Phospho-SirT1 (Ser47) Antibody (green). Actin filaments have been labeled using DY-554 phalloidin (red).
undefined Image 11: Sirtuin Antibody Sampler Kit
Western blot analysis of extracts from control HeLa cells (lane 1) or SirT3 knockout HeLa cells (lane 2) using SirT3 (D22A3) Rabbit mAb #5490 (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the SirT3 knockout hela cells confirms the specificity of the antibody for SirT3.
undefined Image 12: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using SirT1 (D1D7) Rabbit mAb.
undefined Image 13: Sirtuin Antibody Sampler Kit
Western blot analysis of extracts from SirT6 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using SirT6 (D8D12) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). SirT6 WT and KO MEF were kindly provided by Dr. David Lombard, University of Michigan.
undefined Image 14: Sirtuin Antibody Sampler Kit
Flow cytometric analysis of K-562 cells, λ phosphatase treated (blue) or untreated (green), using Phospho-SirT1 (Ser47) Antibody compared to a nonspecific negative control antibody (red).
undefined Image 15: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of SirT6 from HCT 116 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 3) or SirT6 (D8D12) Rabbit mAb (lane 2). Lane 1 is 10% input. Western blot analysis was performed using SirT6 (D8D12) Rabbit mAb.
undefined Image 16: Sirtuin Antibody Sampler Kit
Confocal immunofluorescent analysis of HCT 116 cells, expressing either non-targeting shRNA (shNT) (left) or shSirT6 (right), using SirT6 (D8D12) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
undefined Image 17: Sirtuin Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from mouse liver using SirT3 (D22A3) Rabbit mAb #5490. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 18: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Simple WesternTM analysis of lysates (1.0 mg/mL) from HeLa cells using SirT1 (D1D7) Rabbit mAb #9475. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 19: Sirtuin Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.