Recombinant Anti-SIRT5 antibody [EPR23787-116] (ab259967)

Product name

Anti-SIRT5 antibody [EPR23787-116]
See all SIRT5 primary antibodies
Description

Rabbit monoclonal [EPR23787-116] to SIRT5
Host species

Rabbit
Tested applications

Suitable for: IP, Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- 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.
General notes
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.

Form

Liquid
Storage instructions

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.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR23787-116
Isotype

IgG
Research areas

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

Our Abpromise guarantee covers the use of ab259967 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
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.

Notes
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.

Application notes

Is unsuitable for ICC/IF.

Function

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.
Tissue specificity

Widely expressed.
Sequence similarities

Belongs to the sirtuin family. Class III subfamily.
Contains 1 deacetylase sirtuin-type domain.
Domain

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).
Cellular localization

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
Database links
- 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
Alternative names
- 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

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Datasheet download

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Publishing research using ab259967? Please let us know so that we can cite the reference in this datasheet.

ab259967 has been referenced in 1 publication.

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

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As