LOXL1 Antibody - Azide and BSA Free

Western Blot: LOXL1 Antibody [H00004016-D01P] - Analysis of LOXL1 expression in transfected 293T cell line by LOXL1 polyclonal antibody.Lane 1: LOXL1 transfected lysate(63.10 KDa).Lane 2: Non-transfected lysate.

Immunocytochemistry/Immunofluorescence: LOXL1 Antibody [H00004016-D01P] - Analysis of antibody to LOXL1 on HeLa cell. Antibody concentration 10 ug/ml.

Simple Western: LOXL1 Antibody [H00004016-D01P] - Simple Western lane views show lysates of human Hep3B cell line. Specific bands were detected for LOXL1 using LOXL1 antibody at 1:50 and 1:10 diltutions. This experiment was conducted using the 12-230 kDa separation system. Image from verified customer review.

LOXL1 interacts with apoptosis-related modulators.a Schematic of LOXL1 interactor discovery. The potential interactors were presented at higher levels in the LOXL1 experimental group than in the IgG control group. Two replicates of IP-MS were conducted. b Scatter plots of the log2 ratios of iBAQ intensities for the proteins quantified using MS in the LOXL1 Co-IP samples from two replicates compared with the IgG control samples. Proteins displayed in red displayed a fold change of > 2 in two replicates and were determined to potentially interact with LOXL1. c Protein-protein interactions among potential LOXL1 interactors, as revealed by the STRING database. The gray lines indicate the relationships among these proteins, and the black lines indicate the direct interactions between LOXL1 and other proteins. d LOXL1 interacts with multiple proteins, especially BAG2 in U87 cells overexpressing LOXL1. e Endogenous reciprocal Co-IP assay was further performed to test the protein-protein interaction between LOXL1 and BAG2, BAG3 or HSPA1B in LN18 cells. f BAG2 was depleted by transiently transfecting specific siRNA and siNC was used as a negative control. Indicated antibodies against BAG2 and LOXL1 were used to test their protein levels. The percentages of apoptotic cells were examined. (means +/- SD, unpaired t test, two-tailed). g The scatter plot of correlation data between LOXL1 and BAG2 in glioma was provided by mining TCGA database. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LOXL1 interacts with apoptosis-related modulators.a Schematic of LOXL1 interactor discovery. The potential interactors were presented at higher levels in the LOXL1 experimental group than in the IgG control group. Two replicates of IP-MS were conducted. b Scatter plots of the log2 ratios of iBAQ intensities for the proteins quantified using MS in the LOXL1 Co-IP samples from two replicates compared with the IgG control samples. Proteins displayed in red displayed a fold change of > 2 in two replicates and were determined to potentially interact with LOXL1. c Protein-protein interactions among potential LOXL1 interactors, as revealed by the STRING database. The gray lines indicate the relationships among these proteins, and the black lines indicate the direct interactions between LOXL1 and other proteins. d LOXL1 interacts with multiple proteins, especially BAG2 in U87 cells overexpressing LOXL1. e Endogenous reciprocal Co-IP assay was further performed to test the protein-protein interaction between LOXL1 and BAG2, BAG3 or HSPA1B in LN18 cells. f BAG2 was depleted by transiently transfecting specific siRNA and siNC was used as a negative control. Indicated antibodies against BAG2 and LOXL1 were used to test their protein levels. The percentages of apoptotic cells were examined. (means +/- SD, unpaired t test, two-tailed). g The scatter plot of correlation data between LOXL1 and BAG2 in glioma was provided by mining TCGA database. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LOXL1 regulates BAG2 stability through both its enzymatic activity and direct interaction with BAG2.a, b BAG2 protein levels were increased by LOXL1. U87 cells were transiently transfected with plasmids overexpressing wild type (WT) or ED mutant LOXL1, and Vec was used as a negative control. CHX (cycloheximide, 1 μM) was used to treat cells over time (b). c GST-LOXL1, including full length (FL), 1 to 363 AAs (1-363) and 364 to the end AAs (364-END), was incubated with His-BAG2. d His-BAG2, including full length (FL), 1 to 108 AAs (1-108) and 109 to the end AAs (109-END), was incubated with GST-LOXL1. e Molecular simulations were performed to find the potential sites required for interacting with the BAG domain of BAG2. Then, measurement of LOXL1 enzymatic activity was performed. f D515A mutation reduced the direct interaction between LOXL1 and BAG2. GST-LOXL1 (including WT, N513A and D515A) was incubated with His-BAG2. g The interaction diagram of LOXL1-D515 with BAG2-K186. h U87 cells were transiently transfected with plasmids overexpressing WT, ED or D515A mutant LOXL1. i Endogenous BAG2 was determined with a specific antibody when LOXL1 D515 was mutated into A515. j The reduced interaction between LOXL1 and BAG2 decreased the ability of glioma cells to resist apoptosis. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

VEGFR-Src axis signaling increases LOXL1 expression which positively correlates with BAG2 during glioma progression.a Left panel, inhibitor screen of receptor-mediated signaling pathways required for LOXL1 upregulation in U87 cells; right panel, inhibitor screen of central kinases required for LOXL1 upregulation in U87 cells. b Knocking down of SRC gene by siRNA reduced LOXL1 protein in LN18 cells. c Forced expression of LOXL1 rescued the inhibition of Src kinase activity, as determined using apoptosis assays (means +/- SD, one-way ANOVA). d Upper panel, Promo web software predicted the potential transcriptional factors that bound to the LOXL1 promoter (3000 bp upstream of the TSS); Lower panel, a ChIP assay identified that CEBPA targeted the LOXL1 promoter at 480 base pairs upstream of the TSS in LN18 cells (means +/- SD, unpaired t test, two-tailed). e Knocking down CEBPA reduced LOXL1 expression in LN18 cells. Four pairs of siRNAs were applied to target the CEBPA gene. f Real-time qPCR analyses of glioma specimens were performed. The correlation of LOXL1 expression with CEBPA is shown as R2 and p value. g Representative images of IHC staining of glioma specimens are shown. Scale bar, 100 μm. h Semi-quantitative scoring (using a scale from 0 to 300 points) was conducted, and Pearson’s correlation test was performed and evaluated using the R2 and p value. i Blood LOXL1 levels were measured using ELISA, and 21 glioma specimens were divided into two groups: high (H) or low (L) (means +/- SD, unpaired t test, two-tailed). j BAG2 levels were higher in the H group than in the L group. (means +/- SD, unpaired t test, two-tailed) k Diagram showing the mechanism by which LOXL1 exerts its antiapoptotic activity. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LOXL1 regulates BAG2 stability through both its enzymatic activity and direct interaction with BAG2.a, b BAG2 protein levels were increased by LOXL1. U87 cells were transiently transfected with plasmids overexpressing wild type (WT) or ED mutant LOXL1, and Vec was used as a negative control. CHX (cycloheximide, 1 μM) was used to treat cells over time (b). c GST-LOXL1, including full length (FL), 1 to 363 AAs (1-363) and 364 to the end AAs (364-END), was incubated with His-BAG2. d His-BAG2, including full length (FL), 1 to 108 AAs (1-108) and 109 to the end AAs (109-END), was incubated with GST-LOXL1. e Molecular simulations were performed to find the potential sites required for interacting with the BAG domain of BAG2. Then, measurement of LOXL1 enzymatic activity was performed. f D515A mutation reduced the direct interaction between LOXL1 and BAG2. GST-LOXL1 (including WT, N513A and D515A) was incubated with His-BAG2. g The interaction diagram of LOXL1-D515 with BAG2-K186. h U87 cells were transiently transfected with plasmids overexpressing WT, ED or D515A mutant LOXL1. i Endogenous BAG2 was determined with a specific antibody when LOXL1 D515 was mutated into A515. j The reduced interaction between LOXL1 and BAG2 decreased the ability of glioma cells to resist apoptosis. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

VEGFR-Src axis signaling increases LOXL1 expression which positively correlates with BAG2 during glioma progression.a Left panel, inhibitor screen of receptor-mediated signaling pathways required for LOXL1 upregulation in U87 cells; right panel, inhibitor screen of central kinases required for LOXL1 upregulation in U87 cells. b Knocking down of SRC gene by siRNA reduced LOXL1 protein in LN18 cells. c Forced expression of LOXL1 rescued the inhibition of Src kinase activity, as determined using apoptosis assays (means +/- SD, one-way ANOVA). d Upper panel, Promo web software predicted the potential transcriptional factors that bound to the LOXL1 promoter (3000 bp upstream of the TSS); Lower panel, a ChIP assay identified that CEBPA targeted the LOXL1 promoter at 480 base pairs upstream of the TSS in LN18 cells (means +/- SD, unpaired t test, two-tailed). e Knocking down CEBPA reduced LOXL1 expression in LN18 cells. Four pairs of siRNAs were applied to target the CEBPA gene. f Real-time qPCR analyses of glioma specimens were performed. The correlation of LOXL1 expression with CEBPA is shown as R2 and p value. g Representative images of IHC staining of glioma specimens are shown. Scale bar, 100 μm. h Semi-quantitative scoring (using a scale from 0 to 300 points) was conducted, and Pearson’s correlation test was performed and evaluated using the R2 and p value. i Blood LOXL1 levels were measured using ELISA, and 21 glioma specimens were divided into two groups: high (H) or low (L) (means +/- SD, unpaired t test, two-tailed). j BAG2 levels were higher in the H group than in the L group. (means +/- SD, unpaired t test, two-tailed) k Diagram showing the mechanism by which LOXL1 exerts its antiapoptotic activity. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32424143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Representative immunohistochemistry of LOXs in PDAC tissues and normal pancreas tissues. (A) IHC staining of LOX and quantification showed higher expression in PDAC than normal pancreas tissues. (B) IHC staining and quantification showed higher expression of LOXL1 in PDAC. (C) IHC staining and quantification showed higher expression of LOXL2 in PDAC. (D) IHC staining and quantification showed lower expression of LOXL3 in PDAC. (E) LOXL4 expression levels were similar in PDAC and normal tissues. *p < 0.01; NS, no significant difference. n = 6, repeated 5 times. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35433829), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free

LOXL1 Antibody - Azide and BSA Free Summary

• Description: Quality control test: Antibody reactive against mammalian transfected lysate.
• Immunogen: LOXL1 (NP_005567.2, 1 a.a. - 574 a.a.) full-length human protein. MALARGSRQLGALVWGACLCVLVHGQQAQPGQGSDPARWRQLIQWENNGQVYSLLNSGSEYVPAGPQRSESSSRVLLAGAPQAQQRRSHGSPRRRQAPSLPLPGRVGSDTVRGQARHPFGFGQVPDNWREVAVGDSTGMARARTSVSQQRHGGSASSVSASAFASTYRQQPSYPQQFPYPQAPFVSQYENYDPASRTYDQGFVYYRPAGGGVGAGAAAVASAGVIYPYQPRARYEEYGGGEELPEYPPQGFYPAPERPYVPPPPPPPDGLDRRYSHSLYSEGTPGFEQAYPDPGPEAAQAHGGDPRLGWYPPYANPPPEAYGPPRALEPPYLPVRSSDTPPPGGERNGAQQGRLSVGSVYRPNQNGRGLPDLVPDPNYVQASTYVQRAHLYSLRCAAEEKCLASTAYAPEATDYDVRVLLRFPQRVKNQGTADFLPNRPRHTWEWHSCHQHYHSMDEFSHYDLLDAATGKKVAEGHKASFCLEDSTCDFGNLKRYACTSHTQGLSPGCYDTYNADIDCQWIDITDVQPGNYILKVHVNPKYIVLESDFTNNVVRCNIHYTGRYVSATNCKIVQS
• Specificity: LOXL1 - lysyl oxidase-like 1,
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: LOXL1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Simple Western
  • Western Blot 1:500
• Application Notes: Antibody reactivity against Recombinant Protein with GST tag on ELISA and WB and also on transfected lysate in WB. GST tag alone is used as a negative control. Use in IHC reported in secitific publication PMID: 32424143. LOXL1 Antibody validated for Simple Western from a verified customer review.

Reactivity Notes

Use in Mouse reported in secitific publication PMID: 32424143

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.4)
• Preservative: No Preservative
• Purity: Protein A purified

Notes

This product is produced by and distributed for Abnova, a company based in Taiwan.

Alternate Names for LOXL1 Antibody - Azide and BSA Free

• EC 1.4.3.13
• LOLEC 1.4.3
• LOXL
• LOXL1
• LOXLEC 1.4.3.-
• Lysyl Oxidase Homolog 1
• Lysyl Oxidase like 1
• lysyl oxidase-like 1
• Lysyl oxidase-like protein 1

Background

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. [provided by RefSeq]

Limitations

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.

Publications for LOXL1 Antibody (H00004016-D01P)(6)

Publications using H00004016-D01P

• Jiang C, Wang M, Yao W et al. Comprehensive Analysis on Prognosis and Immune Infiltration of Lysyl Oxidase Family Members in Pancreatic Adenocarcinoma With Experimental Verification Frontiers in Molecular Biosciences 2022-04-01 [PMID: 35433829]
• De Maria A, Zientek KD, David LL et al. Compositional Analysis of Extracellular Aggregates in the Eyes of Patients With Exfoliation Syndrome and Exfoliation Glaucoma Investigative Opthalmology & Visual Science 2021-12-29 [PMID: 34964803]
• Qingjiang H, Takaaki M, Shotaro K et al. Potential association of LOXL1 with peritoneal dissemination in gastric cancer possibly via promotion of EMT. PLoS One. 2020-10-23 [PMID: 33095806]
• Yu H, Ding J, Zhu H et al. LOXL1 confers antiapoptosis and promotes gliomagenesis through stabilizing BAG2 Cell Death Differ. 2020-05-18 [PMID: 32424143] (IF/IHC, WB, Human, Mouse)
• Sethi A, Wordinger RJ, Clark AF. Gremlin utilizes canonical and non-canonical TGFbeta signaling to induce lysyl oxidase (LOX) genes in human trabecular meshwork cells. Exp Eye Res. 2013-08-01 [PMID: 23748100] (WB, Human)
• Sethi A, Mao W, Wordinger RJ, Clark AF. Transforming Growth Factor Beta Induces Extracellular Matrix Protein Crosslinking Lysyl Oxidase (LOX) Genes in Human Trabecular Meshwork Cells. Invest Ophthalmol Vis Sci. 2011-05-05 [PMID: 21546528]

Review for LOXL1 Antibody (H00004016-D01P) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Leah Perkins Simple Western Human 12/06/2022

Summary

• Application: Simple Western
• Sample Tested: hep3b cell line
• Species: Human
• Lot: N/A

Comments

• Comments: ***Bio-Techne Response: This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.*** Test Ab: LOX-L1 Ab Cat#H00004016-D01P, NOVUS Sample: Hep3B cell line Simple protein (Jess): 12-230 kda seperation modules. Best Protein Lysate concentration: 1mg/ml Best Primary Ab dilution: 1:10 Secondary - Rabbit HRP

Bioinformatics

• Gene Symbol: LOXL1
• Entrez:
  • 4016(Human)
• Uniprot:
  • NP_005567.2(Human)