5-Lipoxygenase Antibody - BSA Free

Novus Biologicals | Catalog # NB110-58748

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Knockdown Validated

Cited:

In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all 5-Lipoxygenase Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to a region within residues 100-200 of human 5-Lipoxygenase. [Swiss-Prot# P09917]

Localization

Cytoplasmic

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

80 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for 5-Lipoxygenase Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: 5-Lipoxygenase Antibody - BSA Free [NB110-58748]

Immunocytochemistry/ Immunofluorescence: 5-Lipoxygenase Antibody - BSA Free [NB110-58748]

Immunocytochemistry/Immunofluorescence: 5-Lipoxygenase Antibody - BSA Free [NB110-58748] - U2OS cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with 5-Lipoxygenase Antibody (NB110-58748) at 1ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: 5-Lipoxygenase AntibodyBSA Free [NB110-58748]

Western Blot: 5-Lipoxygenase AntibodyBSA Free [NB110-58748]

Western Blot: 5-Lipoxygenase Antibody [NB110-58748] - Detection of LOX5 in human peripheral leukocytes using NB110-58748 at 0.5(Lane 1) and 2.0 ug/ml (Lane 2).
Simple Western: 5-Lipoxygenase AntibodyBSA Free [NB110-58748]

Simple Western: 5-Lipoxygenase AntibodyBSA Free [NB110-58748]

Simple Western: 5-Lipoxygenase Antibody [NB110-58748] - Simple Western lane view shows a specific band for 5-Lipoxygenase in 0.5 mg/ml of Human Lung lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody
Knockdown Validated: 5-Lipoxygenase Antibody - BSA Free [NB110-58748]

Western Blot: 5-Lipoxygenase Antibody - BSA Free [NB110-58748]

5-Lipoxygenase-Antibody-Knockdown-Validated-NB110-58748-img0007.jpg
5-Lipoxygenase Antibody - BSA Free

Knockdown Validated: 5-Lipoxygenase Antibody - BSA Free [NB110-58748] -

Knockdown of 5-LOX inhibits TNF-alpha -induced IL-6 and MCP-1 release in human synovial fibroblasts.Total protein extracts from RASFs transfected with empty vector or 5-LOX shRNA were used for Western blotting. (A) Immunoblotting showed that cells transfected with 5-LOX shRNA of clone No.3 and No.7 markedly decreased 5-LOX expression. Human synovial fibroblasts transfected with empty vector or 5-LOX shRNA were treated with TNF-alpha for 6 hr. IL-6 and MCP-1 protein release in the conditioned medium was evaluated by ELISA. Note that knockdown of 5-LOX decreased IL-6 (B) and MCP-1 (C) release following TNF-alpha treatment for 6 hr. Data are presented as mean +/- SEM. *, p<0.05 compared with empty vector control (con). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25229347), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for 5-Lipoxygenase Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500 - 1:2000

Simple Western

50 ug/ml

Western Blot

1:500-1:1000
Application Notes
This 5 Lipoxygenase antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot, where a band is seen at approx. 80 kDa. There may be some non-specific bands at non-interfering lower molecular weights. In ICC/IF cytoplasmic staining was observed in Hela cells.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Human Lung lysate 0.5 mg/mL, separated by Size, antibody dilution of 50 ug/mL. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 2 reviews rated 4 using NB110-58748 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: 5-Lipoxygenase

5-lipoxygenase (5LO) is a pro-inflammatory enzyme widely expressed in CNS where its level increases in an age-dependent manner and its activity is regulated by the presence/availability of FLAP (5LO activating protein) that form a functional complex with 5LO to catalyze arachidonic acid oxidation to produce lipid molecules with complex signaling properties such as leukotrienes. Reaction catalyzed by 5LO is considered as first step in the leukotriene biosynthesis and it is highly critical to inflammatory processes. Moreover, knockout of 5LO as well as FLAP has been shown to result in anxiety-like behavior in mice and 5LO is emerging as a new active player in Alzheimer's disease (AD) pathogenesis. 5LO levels are significantly increased in AD postmortem brain tissues and its genetic absence or pharmacological blockade both result in a significant reduction in the abeta levels as well as deposition in a transgenic mouse model of AD-like amyloidosis (i.e., Tg2576) mice.

Alternate Names

5Lipoxygenase, 5LPG, ALOX5, LOG5, LOX5A

Gene Symbol

ALOX5

Additional 5-Lipoxygenase Products

Product Documents for 5-Lipoxygenase Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for 5-Lipoxygenase Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for 5-Lipoxygenase Antibody - BSA Free

Customer Reviews for 5-Lipoxygenase Antibody - BSA Free (2)

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  • 5-Lipoxygenase Antibody
    Name: Bishuang Cai
    Application: Western Blot
    Sample Tested: human macrophage
    Species: Human
    Verified Customer | Posted 06/13/2017
  • 5-Lipoxygenase Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: human macrophage
    Species: Human
    Verified Customer | Posted 05/24/2017

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Protocols

View specific protocols for 5-Lipoxygenase Antibody - BSA Free (NB110-58748):

5-Lipoxygenase Antibody:
Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

5-Lipoxygenase Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 38 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS, 1 hour at room temperature.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-LOX5 primary antibody (NB 110-58748) in blocking buffer and incubate 2 hours at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers
instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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