Recombinant Human Arachidonate 12-lipoxygenase, 12S-type (ALOX12)

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Code CSB-EP001617HU
Size US$306
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Product Details

Purity
Greater than 90% as determined by SDS-PAGE.
Target Names
ALOX12
Uniprot No.
Research Area
Cardiovascular
Alternative Names
12 LOX; 12(S) lipoxygenase ; 12-lipoxygenase; 12LO; 12S LOX; 12S-lipoxygenase; 12S-LOX; 12S-type; Alox12; Arachidonate 12 lipoxygenase; Arachidonate 12 lipoxygenase, 12S type; Arachidonate 12-lipoxygenase; Arachidonate 12-oxidoreductase; Lipoxin synthase 12-LO; LOG12; LOX12_HUMAN; P-12LO; Platelet 12-LOX; Platelet type lipoxygenase 12 ; platelet-type 12-lipoxygenase; Platelet-type lipoxygenase 12
Species
Homo sapiens (Human)
Source
E.coli
Expression Region
1-663aa
Target Protein Sequence
MGRYRIRVATGAWLFSGSYNRVQLWLVGTRGEAELELQLRPARGEEEEFDHDVAEDLGLLQFVRLRKHHWLVDDAWFCDRITVQGPGACAEVAFPCYRWVQGEDILSLPEGTARLPGDNALDMFQKHREKELKDRQQIYCWATWKEGLPLTIAADRKDDLPPNMRFHEEKRLDFEWTLKAGALEMALKRVYTLLSSWNCLEDFDQIFWGQKSALAEKVRQCWQDDELFSYQFLNGANPMLLRRSTSLPSRLVLPSGMEELQAQLEKELQNGSLFEADFILLDGIPANVIRGEKQYLAAPLVMLKMEPNGKLQPMVIQIQPPNPSSPTPTLFLPSDPPLAWLLAKSWVRNSDFQLHEIQYHLLNTHLVAEVIAVATMRCLPGLHPIFKFLIPHIRYTMEINTRARTQLISDGGIFDKAVSTGGGGHVQLLRRAAAQLTYCSLCPPDDLADRGLLGLPGALYAHDALRLWEIIARYVEGIVHLFYQRDDIVKGDPELQAWCREITEVGLCQAQDRGFPVSFQSQSQLCHFLTMCVFTCTAQHAAINQGQLDWYAWVPNAPCTMRMPPPTTKEDVTMATVMGSLPDVRQACLQMAISWHLSRRQPDMVPLGHHKEKYFSGPKPKAVLNQFRTDLEKLEKEITARNEQLDWPYEYLKPSCIENSVTI
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
79.8 kDa
Protein Length
Full Length
Tag Info
N-terminal 6xHis-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
Tris-based buffer,50% glycerol
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

The gene coding for the Human ALOX12 protein (1-663aa) is introduced into a plasmid vector, creating recombinant plasmid that is subsequently transformed into e.coli cells. e.coli cells capable of surviving in the presence of a specific antibiotic are selected and then cultured under conditions favorable for the expression of the gene of interest. A N-terminal 6xHis tag is attached to the protein. Post-expression, the recombinant Human ALOX12 protein is isolated and purified from the cell lysate using affinity purification. Denaturing SDS-PAGE is applied to resolve the resulting recombinant Human ALOX12 protein, revealing a purity exceeding 90%.

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 Q&A
Q:

We ordered your following product CSB-EP001617HU and have the following questions:
1.Need to lyophylized and reconstitute in Tris Buffer. Could you help me with the best protocol for this enzyme?
2. For how long is this recombinant 12-LOX stable in Tris Buffer?

A:
Thanks for your inquiry.
CSB-EP001617HU is shipped with buffer 0.2 μm sterile filtered 10 mM Tris-HCl, 1 mM EDTA, pH 8.0, 50% glycerol. The shipping volume is 250ul, the concentration is 0.2mg/ml.
The buffer contains the glycerol, so if you want to lyophilize, it needs dialysis to remove glycerol first. The protein may loss a lot during the dialysis. The protein that you finally get may less than 50ug.
Can we know why you want to lyophilize and reconstitute in Tris Buffer? If you want the lyophilized protein without glycerol, we suggest you to re-order it. We could offer additional discount.
You also asked how long the protein can be stable in Tris Buffer. At present, we have not done a detailed study on the stability of this. From our current storage situation, the general liquid format protein can keep at -20°C/-80 °C for about 6 months, and it may be longer.
This is related to many factors (such as the state of preservation, buffering composition, temperature of preservation, and stability of the protein itself).
Please feel free to contact me if you need additional information or assistance.

Target Background

Function
Catalyzes the regio and stereo-specific incorporation of molecular oxygen into free and esterified polyunsaturated fatty acids generating lipid hydroperoxides that can be further reduced to the corresponding hydroxy species. Mainly converts arachidonate ((5Z,8Z,11Z,14Z)-eicosatetraenoate) to the specific bioactive lipid (12S)-hydroperoxyeicosatetraenoate/(12S)-HPETE. Through the production of bioactive lipids like (12S)-HPETE it regulates different biological processes including platelet activation. It can also catalyze the epoxidation of double bonds of polyunsaturated fatty acids such as (14S)-hydroperoxy-docosahexaenoate/(14S)-HPDHA resulting in the formation of (13S,14S)-epoxy-DHA. Furthermore, it may participate in the sequential oxidations of DHA ((4Z,7Z,10Z,13Z,16Z,19Z)-docosahexaenoate) to generate specialized pro-resolving mediators (SPMs) like resolvin D5 ((7S,17S)-diHPDHA) and (7S,14S)-diHPDHA, that actively downregulate the immune response and have anti-aggregation properties with platelets. An additional function involves a multistep process by which it transforms leukotriene A4/LTA4 into the bioactive lipids lipoxin A4/LXA4 and lipoxin B4/LXB4, both are vasoactive and LXA4 may regulate neutrophil function via occupancy of specific recognition sites. Can also peroxidize linoleate ((9Z,12Z)-octadecadienoate) to (13S)-hydroperoxyoctadecadienoate/ (13S-HPODE). Due to its role in regulating both the expression of the vascular endothelial growth factor (VEGF, an angiogenic factor involved in the survival and metastasis of solid tumors) and the expression of integrin beta-1 (known to affect tumor cell migration and proliferation), it can be regarded as protumorigenic. Important for cell survival, as it may play a role not only in proliferation but also in the prevention of apoptosis in vascular smooth muscle cells.
Gene References into Functions
  1. ALOX15 orthologs, commonly known as 12/15-lipoxygenases, were suggested to exhibit mainly arachidonic acid 12-lipoxygenating specificity in mammals ranked in evolution lower than gibbons and 15-lipoxygenating specificity in the higher ranking primates [Review]. PMID: 30237084
  2. ALOX12 rs14309 GG genotype expression was found to be significantly associated with Cardiovascular events in patients with Diabetic Nephropathy. PMID: 29196930
  3. Data suggest that ALOX12 is involved in oxidative stress and endoplasmic reticulum stress in liver and other tissues leading to non-alcoholic fatty liver disease. [REVIEW] PMID: 28886991
  4. 12(S)-HETrE, a 12-lipoxygenase oxylipin of dihomo-gamma-linolenic acid, inhibits thrombosis via Galphas signaling in platelets. PMID: 27470510
  5. Results identified a novel ALOX12 locus (indicated by two SNPs in perfect linkage disequilibrium: rs1042357 and rs10852889) that moderated the association between PTSD and reduced thickness of the right prefrontal cortex. PMID: 26372769
  6. The regulation of important oxylipin metabolic genes in peripheral blood mononuclear cells varied with the extent of change in arachidonic acid concentrations in the case of PTGS1 and ALOX12 regulation. PMID: 26672987
  7. ITGB4 stimulation leads to recruitment of 12-LOX from the cytosol to the membrane. PMID: 26037302
  8. In the highest tertile with ALOX12. PMID: 25339205
  9. the amount of LPL expressed in muscle and heart governed both the binding of chylomicron particles and the assimilation of chylomicron lipids in the tissue. PMID: 25598081
  10. Results suggest that Alox12 protein polymorphisms did not discriminate for psoriasis severity. PMID: 24975552
  11. Results indicate that secreted phospholipase A2 IIA (sPLA2-IIA) as an enzyme working in concert with platelet microparticles (MPs) 12-lipoxygenase (12-LO) to promote internalization. PMID: 26106157
  12. IGF-1 reduces lipid oxidation and foam cell formation via downregulation of 12/15-LOX to prevent atherosclerosis. PMID: 25549319
  13. The 12/15-lipoxygenase as an emerging therapeutic target for Alzheimer's disease PMID: 25708815
  14. The costaining of pancreatic polypeptide and vimentin suggests that 12-LO participates in the process leading to beta-cell dedifferentiation in the islet. PMID: 25532042
  15. The ALOX12 levels were relatively decreased in nasal polyp tissue. PMID: 22991985
  16. Inhibition of 12-LOX activity significantly sensitizes prostate cancer cells to radiation. PMID: 25260086
  17. Our goal was to investigate if 12-LOX and/or PAI-1 in patient's plasma could be used to predict outcome of the disease. PMID: 24783193
  18. There was no statistically significant difference in expression of ALOX12 mRNA in cholesteatoma vs. control tissue. PMID: 23832258
  19. This study demonstrated the critical role ALOX12 plays in T. gondii infection in humans, using a monocytic cell line with an ALOX12 knockdown. PMID: 24686056
  20. 12-LOX 261Arg > Gln polymorphism is associated with rectal cancer. PMID: 23715757
  21. PAR4 and GPVI-mediated platelet reactivity involves 12-lipoxygenase PMID: 23784669
  22. The results of our study failed to confirm whether the selected variants in ALOX12 gene within the LT metabolism pathway contribute to platelet reactivity in a diabetic population treated with ASA. PMID: 23828562
  23. Our results suggest that LOX metabolites such as 12-HETE are critical in prostate cancer progression and that the LOX pathway may be a target for treating and preventing prostate cancer. PMID: 22895552
  24. the authors have identified a new pathway by which overexpression of 12-LOX in prostate cancer cells leads to augmented production of MMP9 via activation of PI3K/Akt/NF-kappaB signaling. PMID: 23526143
  25. The LOX isozymes 12 and 15 contribute differentially to striatal vulnerability in response to neurotoxicant challenge. PMID: 23079635
  26. Findings suggest 12/15-LOX expressed in non-epithelial cells such as macrophages and fibroblasts leads to bronchial epithelial injury. PMID: 23528921
  27. Pharmacological inhibition and antisense knockdown of 12LO results in apoptosis of human vascular smooth muscle cells. PMID: 23578768
  28. increased 15-LOX-1 and decreased 5- and 12-LOX levels at the onset of kidney cancer reversing with the progressing stage of the disease or the grade of tumor PMID: 22825379
  29. Non-synonymous polymorphism (Gln261Arg) of 12-lipoxygenase is associated with colorectal and thyroid cancers. PMID: 22864639
  30. results show an increase of ALOX15B during the differentiation of monocytes to macrophages, while the expression of ALOX12 and ALOX15 remains on the same low level. PMID: 22980500
  31. results suggest that genetic variations in arachidonate 12-lipoxygenase (ALOX12) are associated with both the onset and cessation of menstruation in Chinese women living in Shanghai PMID: 22668814
  32. DNA methylation analysis of ALOX12 and GSTM1 in acute myeloid leukaemia identifies prognostically significant groups. PMID: 22924777
  33. The results suggest that the genetic polymorphisms in both human ALOX12 and ALOX15 may contribute to variations in the peak BMD of Chinese women. PMID: 22089472
  34. ALOX12 may have a role in growth of breast cancer, and its inhibition may be a strategy for inhibiting tumor growth PMID: 22384268
  35. DNA hypomethylation associated with persistent wheezing in childhood PMID: 22323304
  36. 12(S)-LOX expression in inflammatory areas of colorectal tumours has the capacity to induce an invasive phenotype in colorectal cancer cells and could be targeted for therapy. PMID: 22237009
  37. Overexpression of 12/15-lipoxygenase leads to increased levels of beta-secretase-1 mRNA and protein, a significant elevation in amyloid beta levels and deposition, and a worsening of memory deficits in Alzheimer's disease transgenic mice. PMID: 22275252
  38. genetic variants in ALOX12 may influence BMD and fracture risk. PMID: 21104233
  39. This study for the first time had co-related the quantity of serum LOX-12 with breast cancer and also with the effect of chemotherapy. PMID: 21945939
  40. 12-LOX may serve as a unique marker and therapeutic target for prostate cancer stem cells PMID: 21225230
  41. the rs2073438 polymorphism of ALOX12 contributes to the variation of obesity phenotypes in young Chinese men PMID: 20697415
  42. Data suggest that the development of combinatory therapies that profit from the ACSL4, lipooxygenase and COX-2 synergistic action may allow for lower medication doses and avoidance of side effects. PMID: 21085606
  43. Protease-activated receptor signaling in platelets activates cytosolic phospholipase A2alpha differently for cyclooxygenase-1 and 12-lipoxygenase catalysis. PMID: 21127289
  44. Genetic absence of this enzyme results in a significant reduction in amyloid-beta (Abeta) production and deposition and an improvement of cognitive deficits. PMID: 20570249
  45. the 12 and 15 ALOX expression, localization and downstream cytokine expression in subcutaneous and omental adipose tissue in human obesity were shown. PMID: 21094135
  46. Thrombin-activated human platelets acutely generate oxidized docosahexaenoic-acid-containing phospholipids via 12-lipoxygenase PMID: 20653566
  47. The mRNA levels of ALOX12 implicated pathways differed significantly in gliomas according to the histological type. PMID: 20382140
  48. we suggest that single nucleotide polymorphisms of the ALOX12 gene might be associated with schizophrenia and negative symptoms in this Korean population PMID: 20626912
  49. Two single-nucleotide polymorphisms, rs9904779 and rs434473 (encodes a replacement of asparagine by serine), of arachidonate 12-lipoxygenase were significantly associated with age at natural menopause PMID: 20061896
  50. ALOX12 is a direct transcriptional target of RUNX1. PMID: 20181616

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Involvement in disease
Esophageal cancer (ESCR); Colorectal cancer (CRC)
Subcellular Location
Cytoplasm, cytosol. Membrane. Note=Membrane association is stimulated by EGF.
Protein Families
Lipoxygenase family
Tissue Specificity
Expressed in vascular smooth muscle cells.
Database Links

HGNC: 429

OMIM: 114500

KEGG: hsa:239

STRING: 9606.ENSP00000251535

UniGene: Hs.654431

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