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Anti-Aspartate Aminotransferase/GOT1 Antibody Picoband™ (monoclonal, 6B3B4)

Figure 1. Western blot analysis of Aspartate Aminotransferase/GOT1 using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
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 30 ug of sample under reducing conditions.
Lane 1: human T-47D whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-Aspartate Aminotransferase/GOT1 antigen affinity purified monoclonal antibody (Catalog # M04085-1) at 0.5 µg/mL overnight at 4°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 # EK1001) with Tanon 5200 system. A specific band was detected for Aspartate Aminotransferase/GOT1 at approximately 43 kDa. The expected band size for Aspartate Aminotransferase/GOT1 is at 46 kDa.
Figure 2. IHC analysis of Aspartate Aminotransferase/GOT1 using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
Aspartate Aminotransferase/GOT1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Aspartate Aminotransferase/GOT1 Antibody (M04085-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 3. IHC analysis of Aspartate Aminotransferase/GOT1 using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
Aspartate Aminotransferase/GOT1 was detected in a paraffin-embedded section of human renal pelvis squamous tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Aspartate Aminotransferase/GOT1 Antibody (M04085-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 4. IHC analysis of Aspartate Aminotransferase/GOT1 using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
Aspartate Aminotransferase/GOT1 was detected in a paraffin-embedded section of human rectum adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Aspartate Aminotransferase/GOT1 Antibody (M04085-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 5. Flow Cytometry analysis of HepG2 cells using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
Overlay histogram showing HepG2 cells stained with M04085-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Aspartate Aminotransferase/GOT1 Antibody (M04085-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 1. Western blot analysis of Aspartate Aminotransferase/GOT1 using anti-Aspartate Aminotransferase/GOT1 antibody (M04085-1).
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 30 ug of sample under reducing conditions.
Lane 1: human T-47D whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-Aspartate Aminotransferase/GOT1 antigen affinity purified monoclonal antibody (Catalog # M04085-1) at 0.5 µg/mL overnight at 4°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 # EK1001) with Tanon 5200 system. A specific band was detected for Aspartate Aminotransferase/GOT1 at approximately 43 kDa. The expected band size for Aspartate Aminotransferase/GOT1 is at 46 kDa.

SPECIFICATION
CITATIONS
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC, FC
Primary Accession	P17174
Host	Mouse
Isotype	IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-Aspartate Aminotransferase/GOT1 Antibody Picoband™ (monoclonal, 6B3B4) . Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	2805
Other Names	Aspartate aminotransferase, cytoplasmic, cAspAT, 2.6.1.1, 2.6.1.3, Cysteine aminotransferase, cytoplasmic, Cysteine transaminase, cytoplasmic, cCAT, Glutamate oxaloacetate transaminase 1, Transaminase A, GOT1 (HGNC:4432)
Calculated MW	43 kDa
Application Details	Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 6B3B4
Immunogen	E.coli-derived human Aspartate Aminotransferase/GOT1 recombinant protein (Position: S5-Q413).
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Protein Information
Name	GOT1 (HGNC:4432)
Function	Biosynthesis of L-glutamate from L-aspartate or L-cysteine (PubMed:21900944). Important regulator of levels of glutamate, the major excitatory neurotransmitter of the vertebrate central nervous system. Acts as a scavenger of glutamate in brain neuroprotection. The aspartate aminotransferase activity is involved in hepatic glucose synthesis during development and in adipocyte glyceroneogenesis. Using L-cysteine as substrate, regulates levels of mercaptopyruvate, an important source of hydrogen sulfide. Mercaptopyruvate is converted into H(2)S via the action of 3-mercaptopyruvate sulfurtransferase (3MST). Hydrogen sulfide is an important synaptic modulator and neuroprotectant in the brain. In addition, catalyzes (2S)-2- aminobutanoate, a by-product in the cysteine biosynthesis pathway (PubMed:27827456).
Cellular Location	Cytoplasm.

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