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FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of SiHa cells using anti- Aldolase/ALDOA antibody (AAA19385).Overlay histogram showing SiHa cells stained with AAA19385 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Aldolase/ALDOA Antibody (AAA19385, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Mouse anti-Human Aldolase/ALDOA Monoclonal Antibody | anti-ALDOA antibody

Anti-Aldolase/ALDOA Antibody (monoclonal, 6H8)

Gene Names
ALDOA; ALDA; GSD12
Reactivity
Human
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay
Purity
Immunogen affinity purified.
Synonyms
Aldolase/ALDOA; Monoclonal Antibody; Anti-Aldolase/ALDOA Antibody (monoclonal; 6H8); Fructose-bisphosphate aldolase A; Lung cancer antigen NY-LU-1; Muscle-type aldolase; ALDOA; ALDA; aldolase; fructose-bisphosphate A; anti-ALDOA antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
Mouse IgG2b
Clone Number
6H8
Specificity
Mouse IgG monoclonal antibody for Aldolase/ALDOA detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-ALDOA antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM)
Application Notes
WB: 0.25-0.5ug/ml|Human|
IHC-P: 2-5ug/ml|Human|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Immunogen
E Coli-derived human Aldolase/ALDOA recombinant protein (Position: E50-Y364).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of SiHa cells using anti- Aldolase/ALDOA antibody (AAA19385).Overlay histogram showing SiHa cells stained with AAA19385 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Aldolase/ALDOA Antibody (AAA19385, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of SiHa cells using anti- Aldolase/ALDOA antibody (AAA19385).Overlay histogram showing SiHa cells stained with AAA19385 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Aldolase/ALDOA Antibody (AAA19385, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 5. IF analysis of Aldolase/ALDOA using anti- Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in immunocytochemical section of HEPG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence) (Figure 5. IF analysis of Aldolase/ALDOA using anti- Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in immunocytochemical section of HEPG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 4. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 3. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 2. IHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Aldolase/ALDOA was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-Aldolase/ALDOA Antibody (AAA19385) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HEPG2 whole cell lysatesLane 2: human A549 whole cell lysatesLane 3: human PC-3whole cell lysatesLane 4: human Hek293 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-Aldolase/ALDOA antigen affinity purified monoclonal antibody (Catalog # AAA19385) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # with Tanon 5200 system. A specific band was detected for Aldolase/ALDOA at approximately 39KD. The expected band size for Aldolase/ALDOA is at 39KD.)

WB (Western Blot) (Figure 1. Western blot analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (AAA19385).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HEPG2 whole cell lysatesLane 2: human A549 whole cell lysatesLane 3: human PC-3whole cell lysatesLane 4: human Hek293 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-Aldolase/ALDOA antigen affinity purified monoclonal antibody (Catalog # AAA19385) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # with Tanon 5200 system. A specific band was detected for Aldolase/ALDOA at approximately 39KD. The expected band size for Aldolase/ALDOA is at 39KD.)
Related Product Information for anti-ALDOA antibody
Aldolase A (ALDOA, or ALDA), also known as fructose-bisphosphate aldolase, is an enzyme that in humans is encoded by the ALDOA gene on chromosome 16. This gene encodes a member of the class I fructose-bisphosphate aldolase protein family. The encoded protein is a glycolytic enzyme that catalyzes the reversible conversion of fructose-1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. Three aldolase isozymes (A, B, and C), encoded by three different genes, are differentially expressed during development. Mutations in this gene have been associated with Glycogen Storage Disease XII, an autosomal recessive disorder associated with hemolytic anemia. Disruption of this gene also plays a role in the progression of multiple types of cancers. Related pseudogenes have been identified on chromosomes 3 and 10.
References
1. Du, S; Guan, Z; Hao, L; Song, Y; Wang, L; Gong, L; Liu, L; Qi, X; Hou, Z; Shao, S (2014). "Fructose-bisphosphate aldolase a is a potential metastasis-associated marker of lung squamous cell carcinoma and promotes lung cell tumorigenesis and migration". PLOS ONE. 9 (1): e85804.
2. Yao, DC; Tolan, DR; Murray, MF; Harris, DJ; Darras, BT; Geva, A; Neufeld, EJ (15 March 2004). "Hemolytic anemia and severe rhabdomyolysis caused by compound heterozygous mutations of the gene for erythrocyte/muscle isozyme of aldolase, ALDOA(Arg303X/Cys338Tyr)". Blood. 103 (6): 2401-3.
3. St-Jean, M; Izard, T; Sygusch, J (11 May 2007). "A hydrophobic pocket in the active site of glycolytic aldolase mediates interactions with Wiskott-Aldrich syndrome protein". The Journal of Biological Chemistry. 282 (19): 14309-15.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
226
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
39,420 Da
NCBI Official Full Name
fructose-bisphosphate aldolase A isoform 1
NCBI Official Synonym Full Names
aldolase A, fructose-bisphosphate
NCBI Official Symbol
ALDOA
NCBI Official Synonym Symbols
ALDA; GSD12
NCBI Protein Information
fructose-bisphosphate aldolase A; muscle-type aldolase; lung cancer antigen NY-LU-1; fructose-1,6-bisphosphate triosephosphate-lyase
UniProt Protein Name
Fructose-bisphosphate aldolase A
UniProt Gene Name
ALDOA
UniProt Synonym Gene Names
ALDA
UniProt Entry Name
ALDOA_HUMAN

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Product Notes

The ALDOA aldoa (Catalog #AAA19385) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-Aldolase/ALDOA Antibody (monoclonal, 6H8) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Aldolase/ALDOA can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM). WB: 0.25-0.5ug/ml|Human| IHC-P: 2-5ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the ALDOA aldoa for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Aldolase/ALDOA, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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