mLrig2-147 | Leucine-rich repeats and immunoglobulin-like domains protein 2
AS14 2788 | Clonality: Polyclonal | Host: Rabbit | Reactivity: mice
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117 | 140 kDa
Thirteen µg of total protein from mouse embryonic fibroblasts (+/+, wild-type; +/-, heterozygous for Lrig2 exon 12; -/-, homozygously deficient of Lrig2 exon 12) extracted with lysis buffer containing 1% Triton X-100 were separated by electrophoresis on a 3 – 8% Tris-acetate NuPAGE gradient gel and blotted to a PVDF membrane. Blots were blocked with 5% nonfat dry milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1,000 over-night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10,000 for 1h at RT with agitation. The blot was washed as above and developed with ECL according to the manufacturer’s instructions.
Reactant: Mus musculus (House mouse)
Application: Western Blotting
Pudmed ID: 24023893
Journal: PLoS One
Figure Number: 1D
Published Date: 2013-09-12
First Author: Rondahl, V., Holmlund, C., et al.
Impact Factor: 2.942Open Publication
Genomic organization of the mouse Lrig2 gene and the generation and molecular analyses of Lrig2-deficient mice.(A) Schematic drawing of the wild-type, conditional, and disrupted Lrig2 alleles. A PKG-neo selection cassette was inserted downstream of exon 12. Exon 12 and the PKG-neo cassette were flanked by loxP sites and were deleted together, in a single step, by mating with OzCre mice. Color coding is as in C. (B) Southern blot using tail DNA isolated from the offspring of an Lrig2E12+/- Ũ Lrig2E12+/- mating. The expected sizes are as follows: wild-type (Lrig2E12+) allele, 11.0 kb; and Lrig2 exon 12-ablated (Lrig2E12-) allele, 6.6 kb. Lane B13 is from Lrig2E12-/-, lanes B10-12 and B14-15 are from Lrig2E12+/-, and the remaining lanes are from Lrig2E12+/+ mice. (C) Genomic organization of the mouse Lrig2 gene. Gene structure of Mus musculus Lrig2 is shown. Gene organization was deduced from the sequence of the mouse genome and the Lrig2 mRNA (GenBank accession number NM_001025067). Exons are indicated with boxes and are drawn to scale. Exon numbers are indicated with numbers. The Lrig2 gene is approximately 58 kb, and it is located on mouse chromosome 3 F2. Color coding depicts the encoded protein domains, including a signal peptide (red), a leucine-rich repeats domain (yellow), three immunoglobulin-like domains (blue), a transmembrane domain (orange), and a cytosolic domain (green). (D) Western blot of MEF cell lines of different Lrig2 genotypes. Top, anti-Lrig2 polyclonal; Bottom, anti-actin. (E) Lrig mRNA levels in mice of different genotypes. The mRNA levels of Lrig1, exon 12-containing Lrig2 (Lrig2E12), exon 17-18 boundary-containing Lrig2 (Lrig2E17-18), and Lrig3 in brains of 3-week old mice were analyzed using quantitative real-time RT-PCR. The Lrig/Rn18s ratio was calculated and normalized to the corresponding ratio in reference RNA from Stratagene. Shown are the means of wild-type (n=6), Lrig2E12+/- (n=5), and Lrig2E12-/- (n=8) mice, with error bars indicating the standard deviations. Significant differences compared with the wild-type mice are indicated with asterisks (*p<0.01 and **p<0.001).
Antibody reactivity was confirmed on mouse embryonic fibrobalsts.
Lrig2 (Leucine-rich repeats and immunoglobulin-like domains protein 2) is coded by Lrig2 gene and belongs to a family of integral membrane proteins. LIRG gene family is composed of three paralogues, LIRG1, LRIG2 and LRIG3.
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