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  1. Link

    lox-antibody-epr4025-ab174316.pdf

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Signal Transduction Cytoskeleton / ECM Extracellular Matrix ECM Enzymes Other Enzymes
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LOX antibody [EPR4025] (ab174316)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (22)

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Flow Cytometry (Intracellular) - Anti-LOX antibody [EPR4025] (ab174316)
  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOX antibody [EPR4025] (ab174316)
  • Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] (ab174316)
  • Immunoprecipitation - Anti-LOX antibody [EPR4025] (ab174316)
  • Anti-LOX antibody [EPR4025] (ab174316)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4025] to LOX
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Primary
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Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Protein
Product image
Recombinant Human LOX protein (Tagged) (ab235570)

View more associated products

Overview

  • Product name

    Anti-LOX antibody [EPR4025]
    See all LOX primary antibodies
  • Description

    Rabbit monoclonal [EPR4025] to LOX
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • HeLa, (HeLa (Human cervix adenocarcinoma epithelial cell) treated with 0.5 mM DFO for 24 hours whole cell lysates ) Jurkat and WI-38 cell lysates. Human muscle tissue. HeLa cells. Immunoprecipitation pellet from WI-38 cells lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4025
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • ECM Enzymes
    • Other Enzymes
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • Other
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Other
    • Cardiovascular
    • Vasculature
    • Endothelium
    • Cardiovascular
    • Vasculature
    • Smooth muscle cell (SMC)
    • Extracellular matrix

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-LOX antibody [EPR4025] (ab197061)
    • Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (ab219369)
    • Anti-LOX antibody [EPR4025] - BSA and Azide free (ab271926)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • WI-38 whole cell lysate (ab3960)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human LOX protein (Tagged) (ab235570)
  • Related Products

    • Anti-LOX antibody [EPR4025] - Low endotoxin, Azide free (ab219369)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab174316 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
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 47 kDa.

For unpurified, use 1/1000 - 1/2000.

IHC-P
1/900. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified, use 1/300.

IHC antigen retrieval protocols.

ICC/IF (1)
1/300.

For unpurified, use 1/100.

IP
1/10 - 1/100.
Flow Cyt (Intra)
1/100.

For unpurified, use 1/30. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 47 kDa.

For unpurified, use 1/1000 - 1/2000.

IHC-P
1/900. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified, use 1/300.

IHC antigen retrieval protocols.

ICC/IF
1/300.

For unpurified, use 1/100.

IP
1/10 - 1/100.
Flow Cyt (Intra)
1/100.

For unpurified, use 1/30. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Responsible for the post-translational oxidative deamination of peptidyl lysine residues in precursors to fibrous collagen and elastin. In addition to cross-linking of extracellular matrix proteins, may have a direct role in tumor suppression.
  • Tissue specificity

    Heart, placenta, skeletal muscle, kidney, lung and pancreas.
  • Involvement in disease

    Defects in LOX may be a cause of cutis laxa autosomal recessive type 1 (ARCL1) [MIM:219100].
  • Sequence similarities

    Belongs to the lysyl oxidase family.
  • Post-translational
    modifications

    The lysine tyrosylquinone cross-link (LTQ) is generated by condensation of the epsilon-amino group of a lysine with a topaquinone produced by oxidation of tyrosine.
  • Cellular localization

    Secreted > extracellular space.
  • Target information above from: UniProt accession P28300 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4015 Human
    • Entrez Gene: 16948 Mouse
    • Entrez Gene: 24914 Rat
    • Omim: 153455 Human
    • SwissProt: P28300 Human
    • SwissProt: P28301 Mouse
    • SwissProt: P16636 Rat
    • Unigene: 102267 Human
    • Unigene: 172 Mouse
    • Unigene: 11372 Rat
    see all
  • Alternative names

    • lox antibody
    • LYOX antibody
    • LYOX_HUMAN antibody
    • Lysyl oxidase antibody
    • MGC105112 antibody
    • Protein lysine 6 oxidase antibody
    • Protein-lysine 6-oxidase antibody
    see all

Images

  • Flow Cytometry (Intracellular) - Anti-LOX antibody [EPR4025] (ab174316)
    Flow Cytometry (Intracellular) - Anti-LOX antibody [EPR4025] (ab174316)

    Flow Cytometry (Intracellular) analysis of ab174316. HeLa (Human cervix adenocarcinoma epithelial cell) cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol. ab174316 was used at 1:100 dilution (1ug) (Red). Secondary antibody Goat anti rabbit IgG (Alexa Fluor® 647, ab150083) at 1:5000 dilution. Isotype control Rabbit monoclonal IgG (ab172730) (Black). Unlabelled control, cells without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    All lanes : Anti-LOX antibody [EPR4025] (ab174316) at 1/1000 dilution

    Lane 1 : Wild-type HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate
    Lane 2 : Wild-type HeLa Treated DFO (0.5 mM, 24 h) cell lysate
    Lane 3 : LOX knockout HeLa Vehicle Control DFO (0 mM, 24 h) cell lysate
    Lane 4 : LOX knockout HeLa Treated DFO (0.5 mM, 24 h) cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-LOX antibody [EPR4025] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab174316 was shown to bind specifically to LOX. A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in Lox knockout cell line ab261801 (knockout cell lysate ab256981). To generate this image, wild-type and Lox knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    All lanes :

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 0.5 mM DFO for 24 hours whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 7 seconds


    Primary antibody dilution:

    ab174316 Anti-LOX antibody 1:5000 dilution (0.1mg/ml)

    ab179483 Anti-HIF-1 alpha antibody 1:500 dilution (0.3mg/ml)

     

    Blocking buffer / Dilution buffer and concentration:

    5% NFDM/TBST

     

    LOX and HIF-1 alpha could be induced under hypoxia condition (PMID: 23545606, PMID: 30226558).

  • Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    Western blot - Anti-LOX antibody [EPR4025] (ab174316)
    All lanes : Anti-LOX antibody [EPR4025] (ab174316) at 1/3700 dilution (purified)

    Lane 1 : WI-38 cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : Mouse brain
    Lane 4 : Rat brain

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOX antibody [EPR4025] (ab174316)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOX antibody [EPR4025] (ab174316)

    Immunohistochemical staining of paraffin embedded human kidney with purified ab174316 at a working dilution of 1 in 900. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

  • Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] (ab174316)
    Immunocytochemistry/ Immunofluorescence - Anti-LOX antibody [EPR4025] (ab174316)

    Immunofluorescence staining of Jurkat cells with purified ab174316 at a working dilution of 1 in 300, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 200. The cells were fixed in 4% PFA.

  • Immunoprecipitation - Anti-LOX antibody [EPR4025] (ab174316)
    Immunoprecipitation - Anti-LOX antibody [EPR4025] (ab174316)

    Western blot analysis on immunoprecipitation pellet from (Lane 1) WI-38 (Human fetal lung fibroblast cell line) cells lysate or (Lane 2) 1X PBS (negative control) using unpurified ab174316 at a 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

  • Anti-LOX antibody [EPR4025] (ab174316)
    Anti-LOX antibody [EPR4025] (ab174316)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (22)

Publishing research using ab174316? Please let us know so that we can cite the reference in this datasheet.

ab174316 has been referenced in 22 publications.

  • Maller O  et al. Tumour-associated macrophages drive stromal cell-dependent collagen crosslinking and stiffening to promote breast cancer aggression. Nat Mater 20:548-559 (2021). PubMed: 33257795
  • Bi X  et al. Collagen Cross-Linking Is Associated With Cardiac Remodeling in Hypertrophic Obstructive Cardiomyopathy. J Am Heart Assoc 10:e017752 (2021). PubMed: 33356379
  • Watanabe K  et al. Group V secreted phospholipase A2 plays a protective role against aortic dissection. J Biol Chem 295:10092-10111 (2020). PubMed: 32482892
  • Despotovic SZ  et al. Altered organization of collagen fibers in the uninvolved human colon mucosa 10?cm and 20?cm away from the malignant tumor. Sci Rep 10:6359 (2020). PubMed: 32286443
  • Klepfish M  et al. LOXL2 Inhibition Paves the Way for Macrophage-Mediated Collagen Degradation in Liver Fibrosis. Front Immunol 11:480 (2020). PubMed: 32296422
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-LOX antibody [EPR4025]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (chondrocytes)
Permeabilization
Yes - Triton x100
Specification
chondrocytes
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 37°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 27 2022

IHC - Wholemount abreview for Anti-LOX antibody [EPR4025]

Average
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Zebrafish Embryo (3 days post fertilization)
Specification
3 days post fertilization
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 22 2017

Western blot abreview for Anti-LOX antibody [EPR4025]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (whole embryo)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
150 µg
Specification
whole embryo
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 25 2017

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