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LOXL2 Recombinant Rabbit Monoclonal Antibody [JU32-53]

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Catalog# ET1706-11

LOXL2 Recombinant Rabbit Monoclonal Antibody [JU32-53]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

  • IF-Tissue

Reactivity

  • Mouse

  • Human

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

LOXL2 Recombinant Rabbit Monoclonal Antibody [JU32-53]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human LOXL2.

Species Reactivity

Mouse, Human, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC, IF-Tissue

Target Molecular Weight

Predicted band size: 87 kDa

Positive Control

HeLa cell lysate,HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate,U-87 MG cell lysate, A431 cell lysate, A431, A549, HUVEC, human placenta tissue, mouse testis tissue, rat brain tissue,A549.

Conjugation

unconjugated

Clone Number

JU32-53

RRID

AB_3070636

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:100

  • FC

  • 1:50-1:100

Target

Function

Lysyl oxidase homolog 2 is an enzyme that in humans is encoded by the LOXL2 gene.This gene encodes a member of the lysyl oxidase gene family. The prototypic member of the family is essential to the biogenesis of connective tissue, encoding an extracellular copper-dependent amine oxidase that catalyses the first step in the formation of crosslinks in collagens and elastin. A highly conserved amino acid sequence at the C-terminus end appears to be sufficient for amine oxidase activity, suggesting that each family member may retain this function. The N-terminus is poorly conserved and may impart additional roles in developmental regulation, senescence, tumor suppression, cell growth control, and chemotaxis to each member of the family.LOXL2 can also crosslink collagen type IV and hence influence the sprouting of new blood vessels. LOXL2 is an enzyme that is up-regulated in several types of cancer and is associated with a poorer prognosis.LOXL2 changes the structure of histones (proteins that are attached to DNA) and thus changes the shape of the cells, making it easier for the cancer cells to metastasize.

Background References

1. Herranz N et al. Lysyl oxidase-like 2 deaminates lysine 4 in histone H3. Mol Cell. 11;46(3):369-76 (2012).

2. Moreno-Bueno G et al. Lysyl oxidase-like 2 (LOXL2), a new regulator of cell polarity required for metastatic dissemination of basal-like breast carcinomas. EMBO Mol Med. 3(9):528-44 (2011).

Sequence Similarity

Belongs to the lysyl oxidase family.

Tissue Specificity

Expressed in many tissues. Highest expression in reproductive tissues, placenta, uterus and prostate. In esophageal epithelium, expressed in the basal, prickle and granular cell layers. Up-regulated in a number of cancers cells and tissues.

Post-translational Modification

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.; N-glycosylated. N-glycosylation on Asn-455 and Asn-644 may be essential for proper folding and secretion; may be composed of a fucosylated carbohydrates attached to a trimannose N-linked glycan core.

Subcellular Location

Secreted, Nucleus.

UNIPROT

SwissProt: Q9Y4K0 Human

SwissProt: P58022 Mouse

SwissProt: B5DF27 Rat

Synonyms

LOR 2 antibody

LOR2 antibody

LOX L2 antibody

LOXL 2 antibody

LOXL2 antibody

LOXL2_HUMAN antibody

Lysyl oxidase homolog 2 antibody

Lysyl oxidase like 2 antibody

Lysyl oxidase like protein 2 antibody

Lysyl oxidase related 2 antibody

Expand

Images

  • Western blot analysis of LOXL2 on different lysates with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate<br />Lane 3: U-87 MG cell lysate<br />Lane 4: MCF7 cell lysate (low expression)<br />Lane 5: A431 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 3 minutes; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 87 kDa<br />Observed band size: 105 kDa

    Western blot analysis of LOXL2 on different lysates with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate
    Lane 3: U-87 MG cell lysate
    Lane 4: MCF7 cell lysate (low expression)
    Lane 5: A431 cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 3 minutes; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1706-11, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 87 kDa
    Observed band size: 105 kDa

  • Immunocytochemistry analysis of A431 cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of A431 cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of A549 cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of A549 cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of HUVEC cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HUVEC cells labeling LOXL2 with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-11) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse placenta tissue with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-11) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-11) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of LOXL2 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of LOXL2 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-11, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-LOXL2 antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-11" style="font-weight: bold;text-decoration: underline;">ET1706-11</a>) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-LOXL2 antibody (ET1706-11) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-11) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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