Anti-Glucose 6 phosphate isomerase antibody (ab86950)
Overview
- Product nameAnti-Glucose 6 phosphate isomerase antibodySee all Glucose 6 phosphate isomerase primary antibodies ...
- DescriptionRabbit polyclonal to Glucose 6 phosphate isomerase
- Tested applicationsICC/IF, IHC-P, WB, IP more details
- Species reactivityReacts with: Rat, Human
Predicted to work with: Mouse, Rabbit, Cow, Macaque Monkey, Orangutan - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Glucose 6 phosphate isomerase.
(Peptide available as ab86951.)
- Positive controlThis antibody gave a positive signal in the following tissue lysates: Rat Heart Rat Lung Human Heart
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab86950 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: Use a concentration of 5 µg/ml. |
| IHC-P | IHC-P: Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).Can be blocked with Glucose 6 phosphate isomerase peptide (ab86951). |
| IP | IP: Use a concentration of 5 µg/ml. |
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
modificationsPhosphorylation 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.
-
Database links
- Entrez Gene: 2821 Human
- Entrez Gene: 14751 Mouse
- Entrez Gene: 292804 Rat
- Omim: 172400 Human
- SwissProt: P06744 Human
- SwissProt: P06745 Mouse
- SwissProt: Q6P6V0 Rat
- Unigene: 466471 Human
- Unigene: 589 Mouse
- Unigene: 84435 Rat
see all
Target information above from: UniProt accession
P06744
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- AMF antibodyAurocrine motility factor antibodyAutocrine motility factor antibody
- DKFZp686C13233 antibodyEC 5.3.1.9 antibodyG6PI_HUMAN antibodyGlucose phosphate isomerase antibodyGlucose-6-phosphate isomerase antibodyGNPI antibodyGPI antibodyGpi1 antibodyHexose monophosphate isomerase antibodyHexosephosphate isomerase antibodyNeuroleukin antibodyNLK antibodyOxoisomerase antibodyPGI antibodyPHI antibodyPhosphoglucose isomerase antibodyPhosphohexomutase antibodyPhosphohexose isomerase antibodyPhosphosaccharomutase antibodySA 36 antibodySA-36 antibodySA36 antibodySperm antigen 36 antibody
see all
Anti-Glucose 6 phosphate isomerase antibody images
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Western blot - Anti-Glucose 6 phosphate isomerase antibody (ab86950)All lanes : Anti-Glucose 6 phosphate isomerase antibody (ab86950) at 1 µg/ml
Lane 1 : Heart (Rat) Tissue Lysate
Lane 2 : Lung (Rat) Tissue Lysate
Lane 3 :Heart (Human) Tissue Lysate - adult normal tissue (ab29431)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 63 kDa
Observed band size : 63 kDa -
Immunocytochemistry/ Immunofluorescence - Glucose 6 phosphate isomerase antibody (ab86950)ICC/IF image of ab86950 stained PC12 cells. The cells were 4% PFA 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 (ab86950, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 5µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Glucose 6 phosphate isomerase antibody (ab86950)IHC image of Glucose 6 phosphate isomerase staining in human colon carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab86950, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Western blot - Anti-Glucose 6 phosphate isomerase antibody (ab86950)All lanes : Anti-Glucose 6 phosphate isomerase antibody (ab86950) at 1 µg/ml
Lane 1 :Glucose 6 phosphate isomerase protein (His tag) (ab87625) at 0.1 µg
Lane 2 :Glucose 6 phosphate isomerase protein (His tag) (ab87625) at 0.01 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 30 seconds -
Immunoprecipitation - Anti-Glucose 6 phosphate isomerase antibody (ab86950)Glucose 6 phosphate isomerase was immunoprecipitated using 0.5mg Rat Heart tissue lysate, 5µg of Rabbit polyclonal to Glucose 6 phosphate isomerase and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat Heart tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab86950.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 63kDa; Glucose 6 phosphate isomerase
References for Anti-Glucose 6 phosphate isomerase antibody (ab86950)
This product has been referenced in:
- Senesi S et al. Contribution of fructose-6-phosphate to glucocorticoid activation in the endoplasmic reticulum: possible implication in the metabolic syndrome. Endocrinology 151:4830-9 (2010). Rat . Read more (PubMed: 20826560) »
