Anti-SIRT5 抗体 [EPR23787-116] (ab259967)

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Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23787-116] to SIRT5
Suitable for: IP, Flow Cyt (Intra), WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

製品名

Anti-SIRT5 antibody [EPR23787-116]
SIRT5 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR23787-116] to SIRT5
由来種

Rabbit
アプリケーション

適用あり: IP, Flow Cyt (Intra), WB, IHC-Pmore details
適用なし: ICC/IF
種交差性

交差種: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: HepG2, Huh7, HeLa, HEK-293T, Hepa1-6, RAW 264.7 and C6 whole cell lysates; Human liver and heart tissue lysates; Mouse liver and heart tissue lysates; Rat liver and tissue lysates. IHC-P: Human liver and ovarian adenocarcinoma tissue; Mouse liver tissue; Rat liver tissue. Flow Cyt (intra): HepG2 and Hepa1-6 cells. IP: HepG2 whole cell lysate; Mouse liver tissue lysate.
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
バッファー

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR23787-116
アイソタイプ

IgG
研究分野

Alternative Versions
- Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab259967の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
IP		1/30.
Flow Cyt (Intra)		1/50.
WB		1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 33 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

特記事項
IP 1/30.
Flow Cyt (Intra) 1/50.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

追加情報

Is unsuitable for ICC/IF.

機能

NAD-dependent lysine demalonylase, desuccinylase and deglutarylase that specifically removes malonyl, succinyl and glutaryl groups on target proteins (PubMed:21908771, PubMed:22076378, PubMed:24703693). Activates CPS1 and contributes to the regulation of blood ammonia levels during prolonged fasting: acts by mediating desuccinylation and deglutarylation of CPS1, thereby increasing CPS1 activity in response to elevated NAD levels during fasting (PubMed:22076378, PubMed:24703693). Activates SOD1 by mediating its desuccinylation, leading to reduced reactive oxygen species (PubMed:24140062). Modulates ketogenesis through the desuccinylation and activation of HMGCS2 (By similarity). Has weak NAD-dependent protein deacetylase activity; however this activity may not be physiologically relevant in vivo. Can deacetylate cytochrome c (CYCS) and a number of other proteins in vitro such as UOX.
組織特異性

Widely expressed.
配列類似性

Belongs to the sirtuin family. Class III subfamily.
Contains 1 deacetylase sirtuin-type domain.
ドメイン

In contrast to class I sirtuins, class III sirtuins have only weak deacetylase activity. Difference in substrate specificity is probably due to a larger hydrophobic pocket with 2 residues (Tyr-102 and Arg-105) that bind to malonylated and succinylated substrates and define the specificity (PubMed:22076378).
細胞内局在

Mitochondrion; Cytoplasm. Mitochondrion and Mitochondrion matrix. Mitochondrion intermembrane space. Cytoplasm, cytosol. Nucleus. Mainly mitochondrial. Also present extramitochondrially: a fraction is present in the cytosol and very small amounts are also detected in the nucleus.
Target information above from: UniProt accession Q9NXA8 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 23408 Human
- Entrez Gene: 68346 Mouse
- Entrez Gene: 306840 Rat
- Omim: 604483 Human
- SwissProt: Q9NXA8 Human
- SwissProt: Q8K2C6 Mouse
- SwissProt: Q68FX9 Rat
- Unigene: 567431 Human
- Unigene: 594133 Human
- Unigene: 35325 Mouse
- Unigene: 137920 Rat
see all
別名
- NAD dependent deacetylase sirtuin 5 antibody
- NAD dependent lysine demalonylase and desuccinylase sirtuin 5 mitochondrial antibody
- NAD dependent protein deacylase sirtuin 5 mitochondrial antibody
- NAD-dependent protein deacylase sirtuin-5, mitochondrial antibody
- Regulatory protein SIR2 homolog 5 antibody
- Silent mating type information regulation 2 S.cerevisiae homolog 5 antibody
- Sir2 like 5 antibody
- SIR2-like protein 5 antibody
- SIR2L5 antibody
- SIR5_HUMAN antibody
- Sirt5 antibody
- Sirtuin 5 antibody
- Sirtuin type 5 antibody
see all

Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

Lane 1 : Mouse liver tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Rat liver tissue lysate
Lane 4 : Rat heart tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 33 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 48 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver (PMID 29491006). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

SIRT5 was immunoprecipitated from 0.35 mg Mouse liver tissue lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse liver tissue lysate 10 ug

Lane 2: ab259967 IP in Mouse liver tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in mouse liver tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate
Lane 6 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 7 : C6 (rat glial tumor glial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 33 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver (PMID 29491006). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

SIRT5 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: ab259967 IP in HepG2 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Flow Cytometry (Intracellular) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Hepa1-6 (Mouse hepatoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

Lane 1 : Human liver tissue lysate
Lane 2 : Human heart tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 33 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver (PMID 29491006). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Flow Cytometry (Intracellular) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Immunohistochemical analysis of paraffin-embedded Human ovarian adenocarcinoma tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human ovarian adenocarcinoma (PMID 31456942). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Anti-SIRT5 antibody [EPR23787-116] (ab259967)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab259967 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab259967 は 3 報の論文で使用されています。

Zheng J et al. CircARF3 Mitigates Allergic Rhinitis through Targeting microRNA-205-5p/Sirtuin 5 Axis. Int Arch Allergy Immunol 184:1056-1070 (2023). PubMed: 37343537
Martino E et al. SIRT3 Modulates Endothelial Mitochondrial Redox State during Insulin Resistance. Antioxidants (Basel) 11:N/A (2022). PubMed: 36009329
Sun J et al. Krüppel-Like Factor 6 Silencing Prevents Oxidative Stress and Neurological Dysfunction Following Intracerebral Hemorrhage via Sirtuin 5/Nrf2/HO-1 Axis. Front Aging Neurosci 13:646729 (2021). PubMed: 34149393

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

ドメイン

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

参考文献 (3)

レビューと Q&A