LRIG1 | Lig-1 (rabbit antibodies)

AS18 4165 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Human

Product Information

Immunogen

KLH-conjugated synthetic peptide from Q96JA1

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS pH 7.4.

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Immunohistochemistry (IHC), Western blot (WB)

Recommended dilution 0.2 µg/ml and 5 µg/ml (IHC), 1:1000 (WB)

Expected | apparent MW

143 kDa

Reactivity

Confirmed reactivity Human

Predicted reactivity

Human, Hominidae,

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Application examples

Application information

Western blot using anti-LRIG-1 antibodies on ovarian cancer

30 µg of total protein from the ovarian cancer cell line Ovsaho was extracted with CEB buffer and EDTA-free protease inhibitor and denatured with Nupage LDS Sample buffer and Nupage Sample reducing agent at 70°C for 10 min. The proteins were separated on Nupage 3-8% Tris-acetate gel and blotted 10 min to PVDF using semi-dry transfer. Blots were blocked with 2 % milk-TBS-T 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 in 2 % milk-TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min (total 30 min) in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit antibody HRP-conjugated) diluted to 1:20 000 in 2 % milk-TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent. Exposure time was 10-20 seconds.

Courtesy Dr. Håkan Hedman, Umeå University, Sweden

IHC using anti-LRIG1 antibodies

Validation of anti-LRIG1 antibody AS184165 for immunohistochemistry. HEK293T cells were rendered LRIG1-null via CRISPR-Cas9 mediated ablation of the entire LRIG1 locus followed by transduction of the cells with a doxycycline-inducible LRIG1 allele encoding FLAG-tagged human LRIG1. LRIG1-expression was induced or not induced followed by pelleting of the cells, paraformaldehyde fixation, paraffin embedment, sectioning, and immunohistochemical staining using an automated Ventana staining machine and different concentrations of anti-LRIG1 or FLAG-M2 antibody. Shown are micrographs of LRIG1-induced cells (left, “Positive control”) and non-induced cells (right, “Negative control”). Anti-LRIG1 antibodies at 5 µg/ml.

Note that the cell population was heterogenous, with only a minority of the induced cells expressing FLAG-LRIG1.

Courtesy Dr. Håkan Hedman, Umeå University, Sweden

Additional information

Peptide used to elicit Lrig1 antibody is not conserved in Lrig3 protein.

IHC was performed on cell pellets made from LRIG1-null HEK293T cells with an inducible LRIG1 allele that had, or had not, been induced to express LRIG1. A specific signal was seen in the antibody range between 0.2 µg/ml and 5 µg/ml.

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Secondary antibodies

Background

The LRIG1 protein Q96JA1 is coded by a gene on chromosome band 3p14.3, a region known to be deleted in various human cancers. This is considered to be a tumour suppressor gene in humans. LRIG1 is an integral cell-surface membrane protein, expressed by specific cells in various human tissues. Its 143-kDa form might be cleaved into 111-kDa and 32-kDa fragments. The LRIG1 protein may inhibit the growth of glial cell tumours and the down-regulation of the LRIG1 gene may be involved in the development and progression of the tumors.

Product citations

Selected references Karlsson et al. (2018). LMO7 and LIMCH1 interact with LRIG proteins in lung cancer, with prognostic implications for early-stage disease. Lung Cancer. 2018 Nov;125:174-184. doi: 10.1016/j.lungcan.2018.09.017.

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