Mouse Legumain ELISA Kit (ab207617)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 8.2 pg/ml
- Range: 54.69 pg/ml - 3500 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
Overview
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Product name
Mouse Legumain ELISA Kit
See all Legumain kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% RAW 264.7 8 3.4% Inter-assay Sample n Mean SD CV% RAW 264.7 3 6% -
Sample type
Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
8.2 pg/ml -
Range
54.69 pg/ml - 3500 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 113 108% - 120% Cell culture media 99 97% - 101% Hep Plasma 113 111% - 115% EDTA Plasma 110 107% - 112% Cit plasma 112 107% - 116% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse -
Product overview
Mouse Legumain ELISA Kit (ab207617) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Legumain protein in cell culture extracts, cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse Legumain with NULL NULL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Legumain is a lysosomal cysteine protease that plays a pivotal role in the endosomal/lysosomal degradation system. Specifically, Legumain is required for the normal degradation of internalized EGFR, allowing Legumain to regulate cell proliferation via this role. Mouse Legumain has 94% and 83% identity to rat and human Legumain, respectively.
Legumain has a strong specificity for hydrolysis of asparaginyl bonds, but can also cleave aspartyl bonds slowly, especially under acidic conditions. Legumain deficiency causes the accumulation of pro-Cathepsins B, H and L, another group of lysosomal cysteine proteases. Overexpression of Legumain in tumors is significant for invasion/metastasis.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Mouse Legumain Capture Antibody 1 x 600µl 10X Mouse Legumain Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Mouse Legumain Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Has a strict specificity for hydrolysis of asparaginyl bonds. Can also cleave aspartyl bonds slowly, especially under acidic conditions. May be involved in the processing of proteins for MHC class II antigen presentation in the lysosomal/endosomal system. -
Tissue specificity
Ubiquitous. Particularly abundant in kidney, heart and placenta. -
Sequence similarities
Belongs to the peptidase C13 family. -
Post-translational
modificationsGlycosylated. -
Cellular localization
Lysosome. - Information by UniProt
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Alternative names
- AEP
- Asparaginyl endopeptidase
- cysteine 1
see all -
Database links
- Entrez Gene: 19141 Mouse
- SwissProt: O89017 Mouse
- Unigene: 17185 Mouse
Images
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Other - Mouse Legumain ELISA Kit (ab207617)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of mouse Legumain standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Example of mouse Legumain standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Legumain in mouse serum and spiked Legumain in RPMI Base Media.The concentrations of Legumain were measured in duplicates, interpolated from the Legumain standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 5%, and RPMI Base Media 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Legumain concentration was determined to be 9.1 ng/mL in serum and 1.7 ng/mL in RPMI Base Media 10%.
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Interpolated concentrations of native Legumain in mouse plasma (heparin, EDTA, and citrate).The concentrations of Legumain were measured in duplicates, interpolated from the Legumain standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (heparin) 5%, plasma (EDTA) 2.5% and plasma (citrate) 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Legumain concentration was determined to be 14.1 ng/mL in plasma (heparin), 7.7 ng/mL in plasma (EDTA) and 6.5 ng/mL in plasma (citrate).
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Interpolated concentrations of native Legumain in mouse RAW 264.7 cell extract based on a 600 µg/mL extract load.The concentrations of Legumain were measured in duplicate and interpolated from the Legumain standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Legumain concentration was determined to be 2.77 ng/mL in mouse RAW 264.7 cell extract.
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Sandwich ELISA - Mouse Legumain ELISA Kit (ab207617)To learn more about the advantages of recombinant antibodies see here.
