Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145]

• WB

Lab

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

purified at 1/3000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg

Lane 4:

Mouse brain lysates at 20 µg

Lane 5:

Mouse heart lysates at 20 µg

Lane 6:

Rat brain lysates at 20 µg

Lane 7:

Rat heart lysates 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

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• WB

Unknown

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

All lanes:

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (ab170950) at 1/1000 dilution

All lanes:

Western blot - Recombinant human FABP-1 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-fabp-1-protein-ab206788'>ab206788</a>) at 0.015 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: 47 kDa

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Exposure time: 180s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) cells labelling Aspartate Aminotransferase + FABP-1 with purified ab170950 at 1/120. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Aspartate Aminotransferase + FABP-1 with purified ab170950 at 1/20 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling Aspartate Aminotransferase + FABP-1 with Purified ab170950 at 1 : 170 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Aspartate Aminotransferase + FABP-1 using unpurified ab170950 at 1/50 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

All lanes:

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (ab170950) at 1/1000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

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• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Immunofluorescent analysis of HepG2 cells labeling Aspartate Aminotransferase + FABP-1 using unpurified ab170950 at 1/50 dilution (green). DAPI nuclear staining (blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Intracellular flow cytometric analysis of permeabilized K562 cells labeling Aspartate Aminotransferase + FABP-1 using unpurified ab170950 at 1/10 dilution (red) or a rabbit IgG negative (green).

• WB

CiteAb

Western blot - Anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] (AB170950)

Aspartate Aminotransferase + FABP-1 western blot using anti-Aspartate Aminotransferase + FABP-1 antibody [EPR12145] ab170950. Publication image and figure legend from Zhu, Y., Li, T., et al., 2017, MBio, PubMed 28811348.

ab170950 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 ab170950 please see the product overview.

GLS2, GLUD1, and GOT2 are upregulated and required for cell proliferation of KSHV-transformed cells. (A) Changes of gene expression of key enzymes in the glutamine metabolism pathway following KSHV transformation. Ratios are untransformed (MM) versus transformed (KMM) cells. Genes downregulated and upregulated by KSHV are labeled in green and red, respectively. (B and C) Analysis of GLS2, GLUD1, GOT1, and GOT2 expression in MM and KMM cells by RT-qPCR (B) and Western blot (C). (D) Examination of the efficiencies of shRNAs against GLS2, GLUD1, and GOT2 in untransformed (MM) and KSHV-transformed (KMM) cells by RT-qPCR. MM and KMM cells infected with lentiviruses harboring 2 different shRNAs (sh1 and sh2) for each gene or a scrambled control (Ctl) were lysed at day 3 postinfection and examined by RT-qPCR. β-Actin was used as an internal control. (E) Examination of the efficiencies of shRNAs against GLS2, GLUD1, and GOT2 in MM and KMM cells by Western blot. Cells were infected as described in the legend for panel D. β-Tubulin was used as an internal control for loading. (F) Knockdown of GLS2, GLUD1, or GOT2 impairs the proliferation of KMM cells. MM and KMM cells infected with lentiviruses harboring 2 different shRNAs for GLS2 (shGLS2-1 and shGLS2-2), GLUD1 (shGLUD1-1 and shGLUD1-2), and GOT2 (shGOT2-1 and shGOT2-2) or a scrambled control (shControl) were counted at different time points after infection.

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