5 Lipoxygenase/5-LO Antibody [G15A12]

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:50

Application
WB, IP, IHC

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
78 kDa

Positive Control	non-Hodgkin's lymphoma; Human RL cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
5 Lipoxygenase/5-LO Antibody [G15A12] detects endogenous levels of total 5 Lipoxygenase/5-LO protein.

Clone
G15A12

Synonym(s)
Polyunsaturated fatty acid 5-lipoxygenase; Arachidonate 5-lipoxygenase (5-LO; 5-lipoxygenase); ALOX5; LOG5

Background
5-Lipoxygenase, also known as 5-LO or ALOX5, is a key enzyme in the lipoxygenase family that catalyzes the initial two-step conversion of arachidonic acid to leukotriene A4, or LTA4, via the intermediate 5-hydroperoxy-eicosatetraenoic acid (5-HPETE). This process requires its nuclear membrane-bound activator FLAP, also called ALOX5AP, which presents the substrate to the enzyme. Human 5-LO exhibits the conserved lipoxygenase fold, featuring an N-terminal beta-sheet domain and a larger alpha-helical catalytic domain that accommodates a non-heme iron, coordinated by key residues His367, His372, His401, Ile417, and Gln558. Its active site features a distinctive, shortened helix alpha2, tilted at approximately 45 degrees, with Phe177 and Tyr181 acting as a "cork" to restrict access; invariant hydrophobic residues such as Leu368, Leu373, Ile406, Leu414, and Leu607 line the elongated U-shaped cavity that positions the pentadiene substrate. 5-LO is primarily expressed in leukocytes, including neutrophils, monocytes, macrophages, and mast cells. In resting cells, it resides in the cytosol or nucleus, but upon stimulation, such as elevated calcium and ATP levels, it translocates to the nuclear membrane, where it associates with FLAP to drive LTA4 synthesis. This results in the production of pro-inflammatory leukotrienes such as LTB4, LTC4, and LTD4, which mediate immune cell chemotaxis, increase vascular permeability, cause bronchoconstriction, and contribute to allergic responses. The activity of 5-LO is tightly regulated by phosphorylation, with kinases like ERK and MAPKAP2 activating the enzyme at Ser271 and Ser663, while PKA inhibits it at Ser523. Additionally, intrinsic instability and auto-inactivation mechanisms ensure that leukotriene production is transient, preventing excessive inflammation. Overexpression of 5-LO is implicated in diseases such as asthma, atherosclerosis, and cancer progression.

Background

5-Lipoxygenase, also known as 5-LO or ALOX5, is a key enzyme in the lipoxygenase family that catalyzes the initial two-step conversion of arachidonic acid to leukotriene A4, or LTA4, via the intermediate 5-hydroperoxy-eicosatetraenoic acid (5-HPETE). This process requires its nuclear membrane-bound activator FLAP, also called ALOX5AP, which presents the substrate to the enzyme. Human 5-LO exhibits the conserved lipoxygenase fold, featuring an N-terminal beta-sheet domain and a larger alpha-helical catalytic domain that accommodates a non-heme iron, coordinated by key residues His367, His372, His401, Ile417, and Gln558. Its active site features a distinctive, shortened helix alpha2, tilted at approximately 45 degrees, with Phe177 and Tyr181 acting as a "cork" to restrict access; invariant hydrophobic residues such as Leu368, Leu373, Ile406, Leu414, and Leu607 line the elongated U-shaped cavity that positions the pentadiene substrate. 5-LO is primarily expressed in leukocytes, including neutrophils, monocytes, macrophages, and mast cells. In resting cells, it resides in the cytosol or nucleus, but upon stimulation, such as elevated calcium and ATP levels, it translocates to the nuclear membrane, where it associates with FLAP to drive LTA4 synthesis. This results in the production of pro-inflammatory leukotrienes such as LTB4, LTC4, and LTD4, which mediate immune cell chemotaxis, increase vascular permeability, cause bronchoconstriction, and contribute to allergic responses. The activity of 5-LO is tightly regulated by phosphorylation, with kinases like ERK and MAPKAP2 activating the enzyme at Ser271 and Ser663, while PKA inhibits it at Ser523. Additionally, intrinsic instability and auto-inactivation mechanisms ensure that leukotriene production is transient, preventing excessive inflammation. Overexpression of 5-LO is implicated in diseases such as asthma, atherosclerosis, and cancer progression.

References
https://pubmed.ncbi.nlm.nih.gov/10591081/ https://pubmed.ncbi.nlm.nih.gov/26427761/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%