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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.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	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	46248 Da
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
Source	Eukaryota
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.

For Research Use Only. Not For Use In Diagnostic Procedures.

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.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Aspartate aminotransferase, cytoplasmic is an enzyme that in humans is encoded by the GOT1 gene. Glutamic-oxaloacetic transaminase is a pyridoxal phosphate-dependent enzyme which exists in cytoplasmic and mitochondrial forms, GOT1 and GOT2, respectively. GOT plays a role in amino acid metabolism and the urea and tricarboxylic acid cycles. The two enzymes are homodimeric and show close homology.

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Cat# ABO16626

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

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