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

Overview

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

    Recombinant GST-tagged fragment (Human)

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

  • FunctionBesides 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.
  • PathwayCarbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 2/4.
  • Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationCytoplasm. 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)

ab66340 has not yet been referenced specifically in any publications.

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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Submitted Aug 07 2012

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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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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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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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Submitted Mar 13 2012

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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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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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Submitted Jul 31 2009

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