Anti-PGAM5 antibody [RM1254] KO Tested (ab322207)

Knockout Tested Rabbit Recombinant Multiclonal PGAM5 antibody. Suitable for WB, IHC-P, ICC/IF, IP and reacts with Human, Mouse, Rat samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (AB322207), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	IP	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Not recommended
Mouse	Tested	Tested	Not recommended	Not recommended	Not recommended
Rat	Tested	Tested	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information PGAM5

Function

Mitochondrial serine/threonine phosphatase that dephosphorylates various substrates and thus plays a role in different biological processes including cellular senescence or mitophagy (PubMed:24746696, PubMed:32439975). Modulates cellular senescence by regulating mitochondrial dynamics. Mechanistically, participates in mitochondrial fission through dephosphorylating DNM1L/DRP1 (PubMed:32439975). Additionally, dephosphorylates MFN2 in a stress-sensitive manner and consequently protects it from ubiquitination and degradation to promote mitochondrial network formation (PubMed:37498743). Regulates mitophagy independent of PARKIN by interacting with and dephosphorylating FUNDC1, which interacts with LC3 (PubMed:24746696). Regulates anti-oxidative response by forming a tertiary complex with KEAP1 and NRF2 (PubMed:18387606). Regulates necroptosis by acting as a RIPK3 target and recruiting the RIPK1-RIPK3-MLKL necrosis 'attack' complex to mitochondria (PubMed:22265414).

Alternative names

Bcl-XL-binding protein v68, Phosphoglycerate mutase family member 5, PGAM5

Recommended products

Knockout Tested Rabbit Recombinant Multiclonal PGAM5 antibody. Suitable for WB, IHC-P, ICC/IF, IP and reacts with Human, Mouse, Rat samples.

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Multiclonal

Immunogens

The exact immunogen used to generate this antibody is proprietary information.

Clone number

RM1254

Purification technique

Affinity purification Protein A

Specificity

Unsuitable for mouse and rat IP.

Unsuitable for mouse and rat ICC.

Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PGAM5 also known as Phosphoglycerate Mutase Family Member 5 is a mitochondrial protein with a known mass of around 32 kDa. It acts as a serine/threonine phosphatase without intrinsic catalytic phosphoglycerate mutase activity. PGAM5 localizes to the inner membrane of mitochondria and exhibits expression in tissues where metabolic regulation is significant. It plays a role in removing phosphate groups from target substrates influencing multiple cellular mechanisms.

Biological function summary

PGAM5 impacts pathways related to cellular stress responses and energy metabolism. It participates in programmed cell death and modulates mitochondrial dynamics. It can serve as part of a complex with other proteins adjusting the balance between mitophagy and apoptosis. Through interaction with key signaling molecules PGAM5 bridges various biochemical processes relevant to cell survival and energy utilization.

Pathways

Its function integrates into mitophagy and necroptosis. PGAM5 collaborates with proteins like PINK1 and PARKIN contributing to the clearance of damaged mitochondria via the mitophagy pathway. In necroptosis PGAM5 interacts with RIPK1 and RIPK3 regulating necrotic cell death. These interactions position PGAM5 at the crossroads of pathways critical to maintaining cellular homeostasis and response to stress.

Associated diseases and disorders

PGAM5 relates to neurodegenerative diseases and cancer. Mutations or dysregulation of PGAM5 connect to Parkinson's disease where impaired mitochondrial function and mitophagy play a role. In cancer altered PGAM5 activity may disrupt normal apoptotic processes leading to unchecked cell growth. In these contexts PGAM5's interaction with proteins like PINK1 PARKIN RIPK1 and RIPK3 underpins its relevance to disease mechanisms.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse testis. The section was incubated with ab322207 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-PGAM5 antibody [RM1254] (ab322207)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, ab322207 was shown to bind specifically to PGAM5. Target of interest was observed at 32 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in PGAM5 knockout cell line (lane 2) (lane 2, knockout cell line Human PGAM5 knockout HeLa cell line ab265141 / knockout cell lysate Human PGAM5 knockout HeLa cell lysate ab257581).
Exposure time: Left panel: 180 seconds; right panel: 70 seconds
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-PGAM5 antibody [RM1254] (ab322207) at 1/1000 dilution
Lane 1: Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: Western blot - Human PGAM5 knockout HeLa cell lysate (Human PGAM5 knockout HeLa cell lysate ab257581) at 20 µg
Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 5: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 32 kDa, 36 kDa
Western blot - Anti-PGAM5 antibody [RM1254] (ab322207)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000. Lanes 3-6 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000.
Exposure time: Lanes 1-4 and 6: 180 seconds; Lane 5: 26 seconds
All lanes: Western blot - Anti-PGAM5 antibody [RM1254] (ab322207) at 1/1000 dilution
Lane 1: Human liver tissue lysate at 20 µg
Lane 2: Human tonsil tissue lysate at 20 µg
Lane 3: Mouse colon tissue lysate at 20 µg
Lane 4: Mouse kidney tissue lysate at 20 µg
Lane 5: Rat colon tissue lysate at 20 µg
Lane 6: Rat liver tissue lysate at 20 µg
Secondary
Lanes 1 - 2: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Lanes 3 - 6: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa
Western blot - Anti-PGAM5 antibody [RM1254] (ab322207)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 22915595 and PMID: 30250224).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-PGAM5 antibody [RM1254] (ab322207) at 1/1000 dilution
Lane 1: Untreated PANC-1(human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: PANC-1 treated with 100 uM CCCP (CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229) for 2 hours, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 36 kDa
Exposure time: 26s
Immunoprecipitation - Anti-PGAM5 antibody [RM1254] (ab322207)
PGAM5 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab322207 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322207 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 2: ab322207 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab322207 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-PGAM5 antibody [RM1254] (ab322207) at 1/30 dilution
All lanes: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 8s
Immunocytochemistry/ Immunofluorescence - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PGAM5 KO HeLa (PGAM5 knockout human cervical adenocarcinoma epithelial cell) (Human PGAM5 knockout HeLa cell line ab265141) cells labelling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).
Confocal image showing mitochondria staining in parental HeLa cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.
Western blot - Anti-PGAM5 antibody [RM1254] (ab322207)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-PGAM5 antibody [RM1254] (ab322207) at 1/1000 dilution
Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: MEF (mouse embryo fibroblast) whole cell lysate at 20 µg
Lane 3: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 36 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat testis. The section was incubated with ab322207 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human colon carcinoma. The section was incubated with ab322207 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human colon. The section was incubated with ab322207 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody [RM1254] (ab322207)
Immunohistochemical analysis of paraffin-embedded A Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet B PGAM5 knockout HeLa cell pellet tissue labeling PGAM5 with ab322207 at 1/100 (4.85 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on (A) wild-type HeLa cell pellet, no staining on (B) PGAM5 knockout HeLa cell pellet. The section was incubated with ab322207 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins