Key features and details
- Mouse monoclonal [1B7D7] to AMF
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-AMF antibody [1B7D7]
See all AMF primary antibodies
DescriptionMouse monoclonal [1B7D7] to AMF
Tested applicationsSuitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
Species reactivityReacts with: Human
Recombinant fragment corresponding to Human AMF.
This product was changed from ascites to supernatant. Lot no’s high than GR185888-22 are from Tissue Culture Supernatant
This product was previously labelled as Glucose 6 phosphate isomerase
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.05% Sodium azide
Concentration information loading...
PurityProtein G purified
Purification notesPurified from TCS.
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
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.
|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.|
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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.
modificationsPhosphorylation at Ser-185 by CK2 has been shown to decrease enzymatic activity and may contribute to secretion by a non-classical secretory pathway.
Cellular localizationCytoplasm. Secreted.
- Information by UniProt
- AMF antibody
- Aurocrine motility factor antibody
- Autocrine motility factor antibody
All lanes : Anti-AMF 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.
All lanes : HRP-conjugated Goat polyclonal to mouse IgG1
Predicted band size: 63 kDa
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.
ab66340 at 1000 dilution staining AMF 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 AMF 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.
ab66340 has been referenced in 12 publications.
- Weng ML et al. Fasting inhibits aerobic glycolysis and proliferation in colorectal cancer via the Fdft1-mediated AKT/mTOR/HIF1a pathway suppression. Nat Commun 11:1869 (2020). PubMed: 32313017
- Fang E et al. Valproic acid suppresses Warburg effect and tumor progression in neuroblastoma. Biochem Biophys Res Commun 508:9-16 (2019). PubMed: 30466785
- Li Y et al. Autocrine motility factor promotes endometrial cancer progression by targeting GPER-1. Cell Commun Signal 17:22 (2019). PubMed: 30836961
- Kwak SE et al. Potential role of exercise-induced glucose-6-phosphate isomerase in skeletal muscle function. J Exerc Nutrition Biochem 23:28-33 (2019). PubMed: 31337203
- Kölbel H et al. Identification of Candidate Protein Markers in Skeletal Muscle of Laminin-211-Deficient CMD Type 1A-Patients. Front Neurol 10:470 (2019). PubMed: 31133972