Anti-Glucose 6 phosphate isomerase antibody [1B7D7] (ab66340)

Overview

  • Product name
    Anti-Glucose 6 phosphate isomerase antibody [1B7D7]
    See all Glucose 6 phosphate isomerase primary antibodies
  • Description
    Mouse monoclonal [1B7D7] to Glucose 6 phosphate isomerase
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, ELISA, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant GST-tagged fragment (Human)

  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR185888-22 are from Tissue Culture Supernatant

Properties

Applications

Our Abpromise guarantee covers the use of ab66340 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/200 - 1/1000.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/500 - 1/5000. Predicted molecular weight: 63 kDa.
ELISA 1/10000.
IP Use at an assay dependent concentration. recommended dilution: 10 ul/mg lysate
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Besides it's role as a glycolytic enzyme, mammalian GPI can function as a tumor-secreted cytokine and an angiogenic factor (AMF) that stimulates endothelial cell motility. GPI is also a neurotrophic factor (Neuroleukin) for spinal and sensory neurons.
  • Pathway
    Carbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 2/4.
  • Involvement in disease
    Defects in GPI are the cause of hemolytic anemia non-spherocytic due to glucose phosphate isomerase deficiency (HA-GPID) [MIM:613470]. It is a form of anemia in which there is no abnormal hemoglobin or spherocytosis. It is caused by glucose phosphate isomerase deficiency. Severe GPI deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment.
  • Sequence similarities
    Belongs to the GPI family.
  • Post-translational
    modifications
    Phosphorylation at Ser-185 by CK2 has been shown to decrease enzymatic activity and may contribute to secretion by a non-classical secretory pathway.
    ISGylated.
  • Cellular localization
    Cytoplasm. Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • AMF antibody
    • Aurocrine motility factor antibody
    • Autocrine motility factor antibody
    • DKFZp686C13233 antibody
    • EC 5.3.1.9 antibody
    • G6PI_HUMAN antibody
    • Glucose phosphate isomerase antibody
    • Glucose-6-phosphate isomerase antibody
    • GNPI antibody
    • GPI antibody
    • Gpi1 antibody
    • Hexose monophosphate isomerase antibody
    • Hexosephosphate isomerase antibody
    • Neuroleukin antibody
    • NLK antibody
    • Oxoisomerase antibody
    • PGI antibody
    • PHI antibody
    • Phosphoglucose isomerase antibody
    • Phosphohexomutase antibody
    • Phosphohexose isomerase antibody
    • Phosphosaccharomutase antibody
    • SA 36 antibody
    • SA-36 antibody
    • SA36 antibody
    • Sperm antigen 36 antibody
    see all

Anti-Glucose 6 phosphate isomerase antibody [1B7D7] images

  • Overlay histogram showing HepG2 cells stained with ab66340 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab66340, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • All lanes : Anti-Glucose 6 phosphate isomerase antibody [1B7D7] (ab66340) at 1/2000 dilution

    Lane 1 : Cell lysates prepared from HepG2 cells.
    Lane 2 : Cell lysates prepared from SMMC-7721 cells.

    Lysates/proteins at 100 µg per lane.

    Secondary
    HRP-conjugated Goat polyclonal to mouse IgG1

    Predicted band size : 63 kDa
  • ab66340 at 1000 dilution staining Glucose 6 phosphate isomerase in L-02 cells by Immunocytochemistry/ Immunofluorescence. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG1 was used as secondary antibody. Green staining in image show positive staining with ab66340, actin filaments were stained red with DY-554 phalloidin and nuclei stained blue with DRAQ5 fluorescent DNA dye.   

  • ab66340 (1µg/ml) staining Glucose 6 phosphate isomerase in human cerebral cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of neurons and of the neuropil.
    Inset panel depicts negative control (no primary antibody).
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References for Anti-Glucose 6 phosphate isomerase antibody [1B7D7] (ab66340)

This product has been referenced in:
  • Xirouchaki CE  et al. Impaired glucose metabolism and exercise capacity with muscle-specific glycogen synthase 1 (gys1) deletion in adult mice. Mol Metab 5:221-32 (2016). Read more (PubMed: 26977394) »
  • Morancho B  et al. Role of ADAM17 in the non-cell autonomous effects of oncogene-induced senescence. Breast Cancer Res 17:106 (2015). Read more (PubMed: 26260680) »

See all 3 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (neuronal)
Total protein in input
500 µg
Specification
neuronal
Immuno-precipitation step
Other - dynabead
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Abcam user community

Verified customer

Submitted Aug 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neuronal)
Specification
neuronal
Fixative
Methanol
Permeabilization
Yes - 0.2% Triton x-100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
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Abcam user community

Verified customer

Submitted Aug 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Purified protein (neuronal)
Loading amount
75 µg
Specification
neuronal
Gel Running Conditions
Non-reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C
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Verified customer

Submitted May 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Mouse Purified protein (neuronal)
Specification
neuronal
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: rt°C
Type
Direct
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Verified customer

Submitted Mar 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human frontal cortex (ICH-P))
Specification
human frontal cortex (ICH-P)
Fixative
Formaldehyde
Permeabilization
Yes - heat-induced in TRIS/EDTA buffer at pH9
Blocking step
gelatine as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
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Verified customer

Submitted May 28 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Huh 7.5 cells)
Loading amount
40 µg
Specification
Huh 7.5 cells
Gel Running Conditions
Reduced Denaturing (12.5% gel)
Blocking step
(agent) for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Verified customer

Submitted Jul 31 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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