Anti-GLO1 antibody (ab129124)
Key features and details
- Rabbit polyclonal to GLO1
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-GLO1 antibody
See all GLO1 primary antibodies -
Description
Rabbit polyclonal to GLO1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human GLO1 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab151614) -
Positive control
- This antibody gave a positive signal in Human Heart and Brain tissue lysates as well as the following whole cell lysates: HepG2; HeLa; HEK293; A431. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129124 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications RatCowChimpanzeeMacaque monkeyOrangutanApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 21 kDa).ICC/IF Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 21 kDa).ICC/IF
Use a concentration of 5 µg/ml.Target
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Function
Catalyzes the conversion of hemimercaptal, formed from methylglyoxal and glutathione, to S-lactoylglutathione. -
Pathway
Secondary metabolite metabolism; methylglyoxal degradation; (R)-lactate from methylglyoxal: step 1/2. -
Sequence similarities
Belongs to the glyoxalase I family. - Information by UniProt
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Database links
- Entrez Gene: 2739 Human
- Entrez Gene: 294320 Rat
- Omim: 138750 Human
- SwissProt: Q04760 Human
- SwissProt: Q6P7Q4 Rat
- Unigene: 268849 Human
- Unigene: 108014 Rat
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Alternative names
- Aldoketomutase antibody
- glo1 antibody
- GLOD1 antibody
see all
Images
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Western blot - Anti-GLO1 antibody (ab129124)All lanes : Anti-GLO1 antibody (ab129124) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GLO1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HepG2 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 21 kDaLanes 1 - 4: Merged signal (red and green). Green - ab129124 observed at 21 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab129124 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in GLO1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GLO1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab129124 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-GLO1 antibody (ab129124)ICC/IF image of ab129124 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab129124 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-GLO1 antibody (ab129124)All lanes : Anti-GLO1 antibody (ab129124) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Human brain tissue lysate - total protein (ab29466)
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 6 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa, 64 kDa, 98 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
The predicted molecular weight of GLO1 is 21 kDa (SwissProt), however we expect to observe a banding pattern around 25 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129124 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab129124 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-GLO1 antibody (ab129124)All lanes : Anti-GLO1 antibody (ab129124) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GLO1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HepG2 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 21 kDaLanes 1 - 4: Merged signal (red and green). Green - ab129124 observed at 21 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab129124 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in GLO1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GLO1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab129124 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-GLO1 antibody (ab129124)
ICC/IF image of ab129124 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab129124 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-GLO1 antibody (ab129124)All lanes : Anti-GLO1 antibody (ab129124) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Human brain tissue lysate - total protein (ab29466)
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 6 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 21 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa, 64 kDa, 98 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
The predicted molecular weight of GLO1 is 21 kDa (SwissProt), however we expect to observe a banding pattern around 25 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129124 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
