- MA1-13029X - Invitrogen Antibodies GLO1 antibody

Submitted references Methylglyoxal-Dependent Glycative Stress and Deregulation of SIRT1 Functional Network in the Ovary of PCOS Mice.

Emidio GD, Placidi M, Rea F, Rossi G, Falone S, Cristiano L, Nottola S, D'Alessandro AM, Amicarelli F, Palmerini MG, Tatone C
Cells 2020 Jan 14;9(1)

Inhibition of Glyoxalase-I Leads to Reduced Proliferation, Migration and Colony Formation, and Enhanced Susceptibility to Sorafenib in Hepatocellular Carcinoma.

Michel M, Hollenbach M, Pohl S, Ripoll C, Zipprich A
Frontiers in oncology 2019;9:785

Glycine Suppresses AGE/RAGE Signaling Pathway and Subsequent Oxidative Stress by Restoring Glo1 Function in the Aorta of Diabetic Rats and in HUVECs.

Wang Z, Zhang J, Chen L, Li J, Zhang H, Guo X
Oxidative medicine and cellular longevity 2019;2019:4628962

Glycine increases glyoxalase-1 function by promoting nuclear factor erythroid 2-related factor 2 translocation into the nucleus of kidney cells of streptozotocin-induced diabetic rats.

Wang Z, Zhao D, Chen L, Li J, Yuan G, Yang G, Zhang H, Guo X, Zhang J
Journal of diabetes investigation 2019 Sep;10(5):1189-1198

Pyruvate kinase M2 activation may protect against the progression of diabetic glomerular pathology and mitochondrial dysfunction.

Qi W, Keenan HA, Li Q, Ishikado A, Kannt A, Sadowski T, Yorek MA, Wu IH, Lockhart S, Coppey LJ, Pfenninger A, Liew CW, Qiang G, Burkart AM, Hastings S, Pober D, Cahill C, Niewczas MA, Israelsen WJ, Tinsley L, Stillman IE, Amenta PS, Feener EP, Vander Heiden MG, Stanton RC, King GL
Nature medicine 2017 Jun;23(6):753-762

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Glyoxylase (Glo1) was performed by loading 25 µg of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a Mouse monoclonal antibody recognizing Glyoxylase (Product # MA1-13029) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a Goat anti-Mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Pico (Product # 34077). For siRNA knock-down experiments HeLa and U20S cells were treated with 100 nM Glo1 siRNA or control siRNA for 48 hours. Lysates were collected and Western blot analysis performed as described above.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), MDA-MB-231 (Lane 3), HL-60 (Lane 4), MOLT-4 (Lane 5), LNCaP (Lane 6), T-47D (Lane 7), U-87 MG (Lane 8) and A549 (Lane 9). The blot was probed with Mouse Anti-GLO1 Monoclonal Antibody (Product # MA1-13029, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 21 kDa band corresponding to GLO1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Glyoxylase (Glo1) was performed by loading 25 µg of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a Mouse monoclonal antibody recognizing Glyoxylase (Product # MA1-13029) at a dilution of 1:1000 overnight at 4C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a Goat anti-Mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Super Signal West Pico (Product # 34077). For siRNA knock-down experiments HeLa and U20S cells were treated with 100 nM Glo1 siRNA or control siRNA for 48 hours. Lysates were collected and Western blot analysis performed as described above.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-GLO1 Monoclonal Antibody (Product # MA1-13029X) and a 21 kDa band corresponding to GLO1 was observed in HeLa, THP-1, HEK-293 and MCF7 cell lines along with Mouse Liver and Rat Liver tissue lysates but not in Mouse Skeletal Muscle tissue lysate which is reported to be negative. Whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), THP-1 (Lane 2), HEK-293 (Lane 3), MCF7 (Lane 4) along with Tissue extracts (30 µg lysate) of Mouse Liver (Lane 5), Mouse Skeletal Muscle (Lane 6) and Rat Liver (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of GLO1 was achieved by transfecting THP-1 with GLO1 specific siRNA (Silencer® select Product # s5823). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the GLO1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with GLO1 Monoclonal Antibody (Product # MA1-13029X, 1 ug/ml) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this Western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GLO1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on cancer biopsies of deparaffinized Human Prostate and Stomach cancer tissues. To expose target proteins high pressure heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer for 20 minutes. Following antigen retrieval endogenous peroxidase activity was quenched with 3% hydrogen peroxide for 10 minutes at room-temp. Tissues were then washed in PBS and blocked in 10% normal goat serum for 20 minutes at room temperature. Tissues were probed at a dilution of 1:800 with a Mouse monoclonal antibody recognizing Glyoxylase (Product # MA1-13029) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBS. Colorimetric detection was performed using metal enhanced DAB and tissues counterstained with hematoxylin. Images are displayed at 40X magnification. Results demonstrate cytoplasmic localization of Glyoxylase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Western blot analysis of glyoxalase 1 (GLO1) ( a ) and GLO2 ( b ) and representative images of immunoreactive bands ( c ) . Data are presented as means +- SEM of densitometric analysis of immunoreactive bands normalized to internal reference protein (glyceraldehyde-3-phosphate dehydrogenase, GAPDH). Three mice per experimental group were employed. Experiments were done in triplicate. ***, p < 0.001, t -test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Expression of Glo-I in human HCC obtained by liver biopsy. (A) Representative image of Glo-I DAB-immunostaining indicates high expression of Glo-I in HCC biopsies. Staining controls lacking the primary antibody are presented in the lower line. (B) Quantification of at least 20 sections per biopsy using the Quick-Score (Q): Q = P [percentage of cells) x I [intensity absent (1), partial (2), complete (3) staining], maximum = 300. Quick-Score was also calculated in non-HCC cirrhotic liver tissue. In all analyzed specimens, Quick-Score of non-HCC tissue was much lower than in HCC tissue (C) Clinical data, including Child-Pugh classes and BCLC stages of analyzed patients.

MA1-13029X

Antibody data