Glyoxalase I Antibody (Glo1a) - BSA Free

Novus Biologicals | Catalog # NBP1-19015

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # Glo1a

Format

BSA Free
View all Glyoxalase I Primary Antibodies »

Product Specifications

Immunogen

Full length human GLO1 protein [Swiss-Prot #Q04760].

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

21 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Glyoxalase I Antibody (Glo1a) - BSA Free

Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015] - Detection of GLO-1 in human cell lines. Lane 1,2 and 3: MG-63 osteosarcoma cell line (positive control), 402-91 liposarcoma cell line and 402-91 liposarcoma cell line resistant to trabectedine. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Immunocytochemistry/ Immunofluorescence: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Immunocytochemistry/Immunofluorescence: Glyoxalase I Antibody (Glo1a) [NBP1-19015] - Immunostaining of human retinal endothelial cells. Immunoreactivity for hGlx-1 was detected in human retinal endothelial cells stained with Glo1a mAb using anti mouse IgG Texas Red as secondary Ab. The cells were counter stained with phalloidin (green) and DAPI (blue), nuclear stain. No reactivity was detected in control cells, stained without Go1a mAb. Figure is a representative of 3 independent experiments. Photo courtesy of Dr. Nagaraj, Case Western Reserve University.
Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Western Blot: Glyoxalase I Antibody (Glo1a) [NBP1-19015] - HeLa whole cell extracts.
Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015] - Simple Western lane view shows a specific band for GLO1 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015]

Simple Western: Glyoxalase I Antibody (Glo1a) [NBP1-19015] - Simple Western lane view shows a specific band detected for GLO1 in HeLa lysate. This experiment was performed under reducing conditions using the Wes or Sally Sue separation system 12-230kDa (or 66-440kDa).

Applications for Glyoxalase I Antibody (Glo1a) - BSA Free

Application
Recommended Usage

ELISA

0.15 ng / 1 ug protein

Immunocytochemistry/ Immunofluorescence

10 ug/ml

Immunohistochemistry-Paraffin

reported in scientific literature (10.3892/ol.2021.12808)

Immunoprecipitation

reported in scientific literature

Simple Western

1:50
Application Notes
This GLO1 antibody is useful for ELISA, Immunocytochemistry/Immunofluorescence and Western blot, where a band is seen approx. 21 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 32 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 1 review rated 5 using NBP1-19015 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

Tris-Glycine and 0.15M NaCl

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: Glyoxalase I

Glyoxalase 1 (GLO1) is the main enzyme in glyoxalase system, a metabolic pathway responsible for detoxifying alpha-oxoaldehydes, particularly methylglyoxal; and is known for converting reactive dicarbonyls into non-toxic intermediates ultimately protecting living cells from producing and accumulating advanced glycation end products, AGEs. GLO1 has also been studied with reference to anxiety, and a murine model showed that this gene's overexpression increases anxiety-related behavior by reducing the GABAA receptor agonist MG. Moreover, an association analysis in a human population with anxiety disorder subtypes suggested a role for GLO1's implication in this phenotype. GLO1 plays an important role in tumour growth and partly in chemotherapy survival as evidenced by a number of studies suggesting that some tumours rely on GLO1 to increase their viability and resistance to multiple chemotherapeutic agents. Furthermore, genetic studies have implicated GLO1 polymorphisms in panic disorder and restless legs syndrome (RLS).

Alternate Names

Aldoketomutase, GLO1, GLOD1, Glx I, GLYI, Methylglyoxalase

Gene Symbol

GLO1

Additional Glyoxalase I Products

Product Documents for Glyoxalase I Antibody (Glo1a) - BSA Free

Certificate of Analysis

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Product Specific Notices for Glyoxalase I Antibody (Glo1a) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Glyoxalase I Antibody (Glo1a) - BSA Free

Customer Reviews for Glyoxalase I Antibody (Glo1a) - BSA Free (1)

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  • Glyoxalase I Antibody (Glo1a)
    Name: Giulia Ribelli
    Application: Western Blot
    Sample Tested: Human osteosarcoma cells – MG63 and Human liposarcoma cells- 402-91
    Species: Human
    Verified Customer | Posted 03/20/2017
    Western Blot: GLO1 Antibody [NBP1-19015] - Detection of GLO-1 in human cell lines. Lane 1,2 and 3: MG-63 osteosarcoma cell line (positive control), 402-91 liposarcoma cell line and 402-91 liposarcoma cell line resistant to trabectedine.
    Glyoxalase I Antibody (Glo1a) - BSA Free NBP1-19015

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Protocols

View specific protocols for Glyoxalase I Antibody (Glo1a) - BSA Free (NBP1-19015):

Glyoxalase I Antibody (Glo1a):
Immunocytochemistry Protocol

1. Cells are cultured one day prior to the experiment.
2. After washing twice with PBS and they are fixed with 4% paraformaldehyde in PBS at ?20C for 15 min.
3. Followed by two washes with PBS, they are permeabilized with 0.1%Triton X-100 in PBS at ?20C for 5 min.
4. To remove the detergent the cells are washed5 times with PBS and then blocked with 2.5 % goat serum in PBS for 2 hr at RT.
5. Cells are then incubated with GLOI mAb (10 μg/ml) in PBS for 1 hr at RT and washed twice for 5 min with PBS.
6. Cells are incubated with secondary antibody (anti-mouseIgG) conjugated with Texas Red (1: 400 dilution in PBS) (Molecular Probes) for 1 hr at RT.

Images of lenses were acquired on a Leica DMI 6000 B inverted microscope using a 20x objective connected to a Retiga EXI camera (Q-imaging Vancouver British Columbia). Secondary Ab contribution to immune reaction was verified by staining without the primary Ab.

Glyoxalase I Antibody (Glo1a):
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-GLO1 primary antibody (NBP1-19015) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted mouse-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, providedit does not interfere with antibody-antigen binding.

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