GPI Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA130853A0HU
Size	US$210
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Image	Western Blot Positive WB detected in: A549 whole cell lysate, U251 whole cell lysate,MCF7 whole cell lysate,HELA whole cell lysate,PC-3 whole cell lysate All lanes: Glucose 6 phosphate isomerase antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 63 kDa Observed band size: 63 kDa IHC image of CSB-RA130853A0HU diluted at 1:50 and staining in paraffin-embedded human prostate tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.45% DAB. IHC image of CSB-RA130853A0HU diluted at 1:50 and staining in paraffin-embedded human lung cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.45% DAB. Overlay Peak curve showing A549 cells stained with CSB-RA130853A0HU (red line) at 1:50. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1µg/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG(H+L) at 1:200 dilution for 35min at 4℃.Control antibody (green line) was rabbit IgG (1µg/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Target Background

Product Details

Uniprot No.

P06744

Target Names

GPI

Alternative Names

Glucose-6-phosphate isomerase (GPI) (EC 5.3.1.9) (Autocrine motility factor) (AMF) (Neuroleukin) (NLK) (Phosphoglucose isomerase) (PGI) (Phosphohexose isomerase) (PHI) (Sperm antigen 36) (SA-36), GPI

Species Reactivity

Human

Immunogen

A synthesized peptide derived from Human GPI

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

12A4

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:2000
IHC	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

In the production of the GPI recombinant monoclonal antibody, in vitro expression systems are employed, involving the cloning of DNA sequences of GPI antibodies obtained from immunoreactive rabbits. The immunogen used is a synthesized peptide derived from the human GPI protein. Subsequently, the genes encoding the GPI antibodies are inserted into plasmid vectors, and these recombinant plasmid vectors are then transfected into host cells to enable antibody expression. Following expression, the GPI recombinant monoclonal antibody undergoes affinity-chromatography purification and is extensively tested for functionality in ELISA, WB, IHC, and FC applications, demonstrating reactivity with the human GPI protein during these assessments.

Glucose-6-phosphate isomerase (GPI) plays a pivotal role in glucose metabolism, particularly in the glycolytic pathway. GPI catalyzes the conversion of glucose-6-phosphate (G6P) to fructose-6-phosphate (F6P), facilitating the breakdown of glucose for energy production and contributing to glucose homeostasis. Additionally, GPI participates in the pentose phosphate pathway, which has roles in nucleotide synthesis and redox balance.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Customer Reviews and Q&A

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Target Background

Function

In the cytoplasm, catalyzes the conversion of glucose-6-phosphate to fructose-6-phosphate, the second step in glycolysis, and the reverse reaction during gluconeogenesis. Besides it's role as a glycolytic enzyme, also acts as a secreted cytokine: acts as an angiogenic factor (AMF) that stimulates endothelial cell motility. Acts as a neurotrophic factor, neuroleukin, for spinal and sensory neurons. It is secreted by lectin-stimulated T-cells and induces immunoglobulin secretion.

Gene References into Functions

Study demonstrated a novel pathway regulating hypoxia-induced angiogenesis in rheumatoid arthritis mediated by glucose-6-phosphate isomerase. PMID: 28067317
Our data suggest that glucose-6-phosphate isomerase gene polymorphism may serve as potential biomarkers to predict the overall survival of hepatocellular carcinoma. PMID: 27288297
Mutation p.Gln413Arg fs*24 is the first frameshift null mutation to be described in GPI deficiency. Molecular modeling suggests that the structural change induced by the p.Gly87Ala pathogenic variant has direct impact in the structural arrangement of the region close to the active site of the enzyme. PMID: 27519939
The 2 cysteines in hGPIc-c are prone to form disulfides upon oxidation. hGPIc-c could induce arthritis in both B10.Q and B10Q.Ncf1*/* mice, whereas hGPIs-s immunization did not induce disease at all, showing that these disulfides are critical for the arthritis-inducing immune response. The cysteines at position 330 and 333 are not crucial for TCR recognition. GILT expression affects hGPI-c-c processing at the CXXC motif. PMID: 29127146
These results suggested that E4P by directly interacting with extracellular HPI/AMF may be an effective strategy to deter BCSC growth and progression. PMID: 28648642
The silencing of PGI/AMF decreased the levels of phosphorylated Akt (-71.9%, P<0.001) compared with the scrambled siRNA, as well as the levels of the stemness marker, SOX2 (-61.7%, P<0.01). Taken together, these findings suggest that PGI/AMF silencing decreases migration, tumorsphere formation as well as the proportion of on the side population cells in glioblastoma U87 cells. PMID: 26936801
NLK is able to promote cell proliferation and type II collagen synthesis during in vitro chondrocyte propagation. PMID: 26573126
Suggest tha AMF/PGI-mediated tumorigenesis occurs through MAPK-ERK signaling in endometrial carcinoma. PMID: 26308071
Study supports a role for AMF mediating epithelial-mesenchymal transition in endometrial cancer (EC) through MAPK signaling. Therefore, AMF may provide a potential prognostic and therapeutic target in preventing EC progression. PMID: 26201353
Results showed an increase in GPI and AMFR in clear cell-renal cell carcinoma cells, and their colocalization on plasma membrane. Kaplan-Meier curves showed significant differences in survival among groups of patients with high versus low GPI. PMID: 26579829
The association of ENO1 and GPI with postthaw sperm viability and motility was confirmed using Pearson's linear correlation. ENO1 and GPI can be used as markers of human sperm freezability before starting the cryopreservation procedure. PMID: 25910678
High GPI expression is associated with metastatic phenotype of breast cancer. PMID: 24440856
PGI is a moonlighting protein that functions as a cytosolic enzyme involved in glycolysis and gluconeogenesis, and as cytokine through binding to its cell surface receptor. PMID: 11004567
Serum anti-GPI autoantibodies are useful for the diagnosis of rheumatoid arthritis in Chinese patients. PMID: 23773638
Findings suggest that autocrine motility factor (AMF)-HER2 interaction might be a novel target for therapeutic management of patients with breast cancer, whose disease is resistant to trastuzumab. PMID: 23248119
For PGI, an extended active site in which residues in the first, second, and third layers around the reacting substrate are predicted. PMID: 21970785
GPI is a target gene of the BACH1 transcription factor according to ChIP-seq analysis in HEK 293 cells. PMID: 21555518
By regulating ER calcium release, AMF/PGI interaction with gp78/AMFR therefore protects against ER stress identifying novel roles for these cancer-associated proteins in promoting tumor cell survival. PMID: 21252914
Data suggest that elevated serum glucose-6-phosphate isomerase may be involved in the synovitis of rheumatoid arthritis and may prove useful as a serum marker for disease activity. PMID: 20810510
Data show that effective downregulation of AMF/PGI expression and subsequent abrogation of AMF/PGI secretion, which resulted in morphologic change with reduced growth, motility, and invasion. PMID: 20978190
autocrine motility factor/phosphoglucose isomerase against TGF-beta-induced apoptosis was correlated with its enzymatic activity PMID: 19819066
hypoxia-induced gene in pancreatic cancer cell lines PMID: 11688991
Overexpression induces neoplastic transformation and survival of NIH-3T3 fibroblasts. PMID: 12517804
The results demonstrate that the enzymatic activity of PGI is not essential for either receptor binding or cytokine function of human PGI. PMID: 12527360
crystal structure and analysis of the initial ring-opening step of catalysis PMID: 12573240
role in regulating cell proliferation PMID: 12783864
AMF regulates expression of Apaf-1 and caspase-9 genes via a complex signaling pathway and indirectly regulates formation of the apoptosome. (autocrine motility factor) PMID: 14566819
The observations are consistent with a downstream mediation role of MMP-3 in Pohophoglucose isomerase/AMF-stimulated tumor cell metastasis. PMID: 14715248
T-cell dependent peripheral polyarthritis induced by recombinant human glucose-6-phosphate isomerase in genetically unaltered mice demonstrates for the first time the induction of organ-specific disease by systemic autoimmunity. PMID: 15034067
In addition to the findings in rheumatoid arthritis, our results indicate that GPI is not a general target of autoantibodies in juvenile idiopathic arthritis. PMID: 15290745
Our results suggest that GPI variants may play a crucial role in the production of autoantibodies against ubiquitous GPI autoantigens. PMID: 15369782
AMF expression significantly contributes to the aggressive phenotype of pancreatic cancer PMID: 15570012
phosphoglucose isomerase/autocrine motility factor activities are differentially regulated by protein kinase CK2 phosphorylation PMID: 15637053
interaction with hypoxia-inducible factor-1 drives mobility of erythroid progenitor cells PMID: 15850830
N-glyco side-chain of AMFR is a trigger and that interaction between the 117-C-terminal part of AMF and the extracellular core protein of autocrine motility factor receptor (AMFR) is needed during AMF-AMFR interactions PMID: 16563432
elevated G6PI levels present in patients with immune-based inflammatory arthritis may contribute to elevated levels of anti-G6PI Abs and G6PI/anti-G6PI immune complexes PMID: 16949042
Missense mutations c.341A>T (p.Asp113Val) in exon 4 and c.663T>G (p.Asn220Lys) in exon 7 are associated with hereditary nonspherocytic hemolytic anemia. PMID: 17041899
the receptor molecule for AMF/NLK/MF in leukemic differentiation is not gp78 PMID: 17071500
raft-dependent endocytosis of AMF follows a distinct phosphatidylinositol 3-kinase-dependent pathway that is up-regulated in more aggressive tumor cells PMID: 17690101
PGI/AMF is involved in oxidative stress-induced cellular senescence and should bring novel insights into the control of cellular growth leading to a new methodology for cancer treatment. PMID: 17925402
IL-6 and Th17 play an essential role in GPI-induced arthritis PMID: 18311788
Results of this study suggest that AMF stimulation stimulates MMP3 expression via a MAPK signaling pathway. PMID: 18485900
Peptide fragment glucose phosphate isomerase (GPI)325-339 is identified as a major epitope in GPI-induced arthritis, with potential to induce polyarthritis. PMID: 18992137
Mutations effect catalytic activity and structural stability of human glucose-6-phosphate isomerase. PMID: 19064002
Expression of this protein leads to mesenchymal-topepithelial transition in breast cancer cells. PMID: 19531650
overexpression of PGI significantly contributes to the aggressive phenotype of human colon cancer PMID: 19787266
melanoma migration induced by AMF is mediated by autocrine production of IL-8 as a novel downstream modulator of the AMF signaling pathway PMID: 19801670

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Involvement in disease

Hemolytic anemia, non-spherocytic, due to glucose phosphate isomerase deficiency (HA-GPID)

Subcellular Location

Cytoplasm. Secreted.

Protein Families

GPI family

Database Links

HGNC: 4458

OMIM: 172400

KEGG: hsa:2821

UniGene: Hs.466471

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