Anti-Aldolase antibody [EPR23181-39] (ab252953)

Rabbit Recombinant Monoclonal Aldolase antibody. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples. Cited in 5 publications.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-Aldolase antibody [EPR23181-39] (AB252953), expandable thumbnail

Publications

Key facts

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

Immunogen

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

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

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

Tested
Tested

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes -
Species Human	Dilution info 1/50	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
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.
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.

Target data

See full target information ALDOA

Function

Catalyzes the reversible conversion of beta-D-fructose 1,6-bisphosphate (FBP) into two triose phosphate and plays a key role in glycolysis and gluconeogenesis (PubMed:14766013). In addition, may also function as scaffolding protein (By similarity).

Alternative names

ALDA, ALDOA, Fructose-bisphosphate aldolase A, Lung cancer antigen NY-LU-1, Muscle-type aldolase

Recommended products

Rabbit Recombinant Monoclonal Aldolase antibody. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples. Cited in 5 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23181-39
Purification technique: Affinity purification Protein A
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

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

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

Activity summary

Aldolase also known as fructose-bisphosphate aldolase is an enzyme that plays a critical role in glycolysis catalyzing the reversible cleavage of fructose 16-bisphosphate into glyceraldehyde-3-phosphate and dihydroxyacetone phosphate. Aldolase is a homotetramer with a molecular mass of approximately 158 kDa. It is highly expressed in liver muscle and brain tissues. Aldolase consists of three isoforms: aldolase A B and C each predominant in different tissues contributing to tissue-specific roles within the organism.

Biological function summary

Aldolase catalyzes an important reaction in energy metabolism ensuring the continuation of glycolytic flux which is essential for ATP production. Aldolases do not form part of larger protein complexes. However they interact with other enzymes within the glycolytic pathway to facilitate a smooth metabolic flow. Moreover aldolase A plays an additional role in gluconeogenesis the pathway involved in synthesizing glucose from non-carbohydrate sources.

Pathways

Aldolase is an essential part of glycolysis and gluconeogenesis pathways. In glycolysis it partners with enzymes like phosphofructokinase and enolase to breakdown glucose for energy production. During gluconeogenesis aldolase works with enzymes including fructose-16-bisphosphatase to create glucose vital for maintaining blood sugar levels during fasting. Through these pathways aldolase ensures balance between energy-producing and energy-consuming states within the cell.

Associated diseases and disorders

Mutations or deficiencies in aldolase particularly aldolase A have been linked to glycogen storage disease type XII which results in muscle weakness and exercise intolerance. Aldolase B deficiency is known to cause hereditary fructose intolerance leading to liver and renal complications. These conditions highlight the importance of aldolase in maintaining metabolic health with implications for proteins within the same pathways potentially affecting their functions and expression levels.

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.

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15 product images

Immunocytochemistry/ Immunofluorescence - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 cells labelling Aldolase with ab252953 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing strong cytoplasmic and weak nuclear staining in A549 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution.
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: Lanes 1-6: 5.5 seconds; Lanes 7-8: 26 seconds.
All lanes: Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953) at 1/1000 dilution
Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 3: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 40 µg
Lane 4: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 40 µg
Lane 5: C2C12 (mouse myoblasts myoblast) whole cell lysate at 40 µg
Lane 6: C6 (rat glial tumor glial cell) whole cell lysate at 40 µg
Lane 7: A549 (human lu carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 8: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 40 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 39 kDa
Observed band size: 39 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Aldolase with ab252953 at 1/50 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining in rat skeletal muscle (PMID: 18676612). The section was incubated with ab252953 for 20 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry (Intracellular) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized A549 (Human lung carcinoma epithelial cell) cells labelling Aldolase with ab252953 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: Lane 1: 10 seconds; Lane 2: 3 seconds.
All lanes: Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953) at 1/1000 dilution
Lane 1: Rat brain tissue lysate at 20 µg
Lane 2: Rat skeletal muscle tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 39 kDa
Observed band size: 39 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Aldolase with ab252953 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing strong cytoplasmic and weak nuclear staining in NIH/3T3 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Aldolase with ab252953 at 1/50 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining in mouse skeletal muscle (PMID: 18676612). The section was incubated with ab252953 for 20 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: Lanes 1-3: 3 seconds; Lane 4-5: 5.5 seconds.
All lanes: Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953) at 1/1000 dilution
Lane 1: Human brain tissue lysate at 40 µg
Lane 2: Human skeletal muscle tissue lysate at 40 µg
Lane 3: Mouse brain tissue lysate at 40 µg
Lane 4: Mouse skeletal muscle tissue lysate at 40 µg
Lane 5: Human colon cancer tissue lysate at 40 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 39 kDa
Observed band size: 39 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Aldolase with ab252953 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining in human colon cancer (PMID: 29453983). The section was incubated with ab252953 for 20 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Immunohistochemical analysis of paraffin-embedded Human lung cancer (A) and its adjacent noncancerous tissue (B) tissue labeling Aldolase with ab252953 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining in human lung cancer (A) while weak or negative staining in its adjacent noncancerous tissue (B) (PMID: 24465716). The section was incubated with ab252953 for 20 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry (Intracellular) - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Aldolase with ab252953 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
The identities of the higher MW bands between 50 and 70 kDa are unknown.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution, anti-Aldolase antibody (ab252953) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Aldolase A / B/ C (lactyl K147) antibody [EPR24696-317] (Anti-Aldolase A / B/ C (lactyl K147) antibody [EPR24696-317] ab313325) at 1/1000 dilution
Lane 1: Untreated C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 2: C6 treated with 100nM Rotenone for 24h, 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 1/20000 dilution
Developed using the ECL technique.
Observed band size: 39 kDa
Exposure time: 180s
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution, anti-Aldolase antibody (ab252953) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Aldolase A / B/ C (lactyl K147) antibody [EPR24696-317] (Anti-Aldolase A / B/ C (lactyl K147) antibody [EPR24696-317] ab313325) at 1/1000 dilution
Lane 1: Untreated NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: NCI-H1299 treated with 100nM Rotenone for 24h, 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 1/20000 dilution
Developed using the ECL technique.
Observed band size: 39 kDa
Exposure time: 59s
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Western blot: Anti-ALDOA antibody [EPR23181-39] (ab252953) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab252953 was shown to bind specifically to ALDOA. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953) at 1/1000 dilution
Lane 1: HEK-293T overexpressing ALDOA cell lysate at 20 µg
Lane 2: HEK-293T control cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Lane 5: Human heart cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 45 kDa
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-Aldolase antibody [EPR23181-39] (ab252953)
Aldolase western blot using anti-Aldolase antibody [EPR23181-39] ab252953. Publication image and figure legend from Lin, J. X., Lian, N. Z., et al., 2022, Cell Death Dis, PubMed 35568711.

ab252953 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab252953 please see the product overview.
LHPP suppresses aerobic glycolysis.A Relative mRNA expression levels of glycolytic genes and glutamine transporters in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells by qPCR. B Protein expression levels of glycolytic genes and glutamine transporters in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells on western blot. C IHC staining of LHPP and HIF1A in TMAs and their correlation. Scale bars = 100 μm. D Oxygen consumption rate and extracellular acidification rate of LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells were measured using the Seahorse Bioscience XF96 analyser. E Human phosphokinase microarray assay analysis of the conditioned medium from stably transfected HGC-27 cells. A summary of the relative signal intensities of the indicated proteins is shown. G, H IHC staining of LHPP and p-GSK3b in TMAs and their correlation. Scale bars = 200 μm. F Combined LHPP and GSK3b by co-immunoprecipitation. I Protein expression levels of the Wnt pathway in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells on western blot. Data were analyzed using the Wilcoxon test. *P < 0.05, **P < 0.01, ***P < 0.001, KEGG Kyoto encyclopaedia of genes and genomes, GO gene ontology, qPCR quantitative polymerase chain reaction, IHC immunohistochemical, TMA tissue microarray.