Anti-AMF antibody [1B7D7] (ab66340)
![Western blot - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.com/ps/products/66/ab66340/Images/ab66340-454787-anti-amf-antibody-1b7d7-western-blot-wildtype-hek293t-gpi-knockout-hek293t.jpg "Western blot - Anti-AMF antibody [1B7D7] (ab66340)")
Key features and details
- Mouse monoclonal [1B7D7] to AMF
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-AMF antibody [1B7D7]
See all AMF primary antibodies -
Description
Mouse monoclonal [1B7D7] to AMF -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human AMF.
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Positive control
- WB: HEK293T, HepG2 and SMMC-7721 cell lysate. Flow Cyt: HepG2 cells. ICC: L-02 cells. IHC-P: Human cerebral cortex tissue.
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General notes
This product was changed from ascites to supernatant. Lot no’s high than GR185888-22 are from Tissue Culture Supernatant.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Purified from TCS. -
Clonality
Monoclonal -
Clone number
1B7D7 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
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) |
1/200 - 1/1000.
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| IHC-P | (1) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| WB | (2) |
1/500 - 1/5000. Predicted molecular weight: 63 kDa.
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| Flow Cyt |
1/100.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
ICC/IF
1/200 - 1/1000. |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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WB
1/500 - 1/5000. Predicted molecular weight: 63 kDa. |
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Flow Cyt
1/100. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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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
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 localization
Cytoplasm. Secreted. - Information by UniProt
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Database links
- Entrez Gene: 2821 Human
- Omim: 172400 Human
- SwissProt: P06744 Human
- Unigene: 466471 Human
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Alternative names
- AMF antibody
- Aurocrine motility factor antibody
- Autocrine motility factor antibody
see all
Images
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Western blot - Anti-AMF antibody [1B7D7] (ab66340)All lanes : Anti-AMF antibody [1B7D7] (ab66340) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : GPI knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 63 kDaLanes 1 - 2: Merged signal (red and green). Green - ab66340 observed at 63 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab66340 was shown to react with AMF in wild-type HEK-293T cells in Western blot with loss of signal observed in GPI knockout cell line ab266834 (GPI knockout cell lysate ab257458). Wild-type HEK-293T and GPI knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab66340 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Flow Cytometry - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.com/ps/products/66/ab66340/Images/ab66340-195306-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-flow-cytometry.jpg)
Flow Cytometry - Anti-AMF antibody [1B7D7] (ab66340)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. -
![Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.com/ps/products/66/ab66340/Images/ab66340-80124-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [1B7D7] (ab66340)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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.com/ps/products/66/ab66340/Images/ab66340-80123-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [1B7D7] (ab66340)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. -
![Western blot - Anti-AMF antibody [1B7D7] (ab66340)](https://www.abcam.com/ps/products/66/ab66340/Images/ab66340-80125-anti-glucose-6-phosphate-isomerase-antibody-1b7d7-western-blot.jpg)
Western blot - Anti-AMF antibody [1B7D7] (ab66340)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.
Secondary
All lanes : HRP-conjugated Goat polyclonal to mouse IgG1
Predicted band size: 63 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (16)
ab66340 has been referenced in 16 publications.
- Zhao H et al. LINC01816 promotes the migration, invasion and epithelial-mesenchymal transition of thyroid carcinoma cells by sponging miR-34c-5p and regulating CRABP2 expression levels. Oncol Rep 45:N/A (2021). PubMed: 33786631
- 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
- Tanie Y et al. GRP78-Mediated Signaling Contributes to Axonal Growth Resulting in Motor Function Recovery in Spinal Cord-Injured Mice. Front Pharmacol 11:789 (2020). PubMed: 32547396
- Sun RC et al. Nuclear Glycogenolysis Modulates Histone Acetylation in Human Non-Small Cell Lung Cancers. Cell Metab 30:903-916.e7 (2019). PubMed: 31523006
- Li H et al. Therapeutic targeting of circ-CUX1/EWSR1/MAZ axis inhibits glycolysis and neuroblastoma progression. EMBO Mol Med 11:e10835 (2019). PubMed: 31709724