Anti-Aldolase antibody [EPR23181-39]

• Flow Cyt (Intra)

Unknown

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 (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.

• ICC/IF

Unknown

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 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. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution.

• IHC-P

Unknown

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 (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 (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

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 (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 (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

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 (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 (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

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 (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 (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Unknown

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 (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.

• ICC/IF

Unknown

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 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. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution.

• WB

Lab

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] (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

Observed band size: 45 kDa

false

• WB

Lab

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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

Lab

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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

Supplier Data

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 (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] (<a href='/en-us/products/primary-antibodies/aldolase-a-b-c-lactyl-k147-antibody-epr24696-317-ab313325'>ab313325</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 39 kDa

true

Exposure time: 59s

• WB

Lab

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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

Supplier Data

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 (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] (<a href='/en-us/products/primary-antibodies/aldolase-a-b-c-lactyl-k147-antibody-epr24696-317-ab313325'>ab313325</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 39 kDa

true

Exposure time: 180s

• WB

CiteAb

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.

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