Rabbit Anti-Legumain antibody

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AEP; Asparaginyl endopeptidase; EC 3.4.22.34; LGMN; LGMN1; Protease, cysteine 1; protease, cysteine, 1 (legumain); PRSC1; LGMN_HUMAN.

Cat:

SL3907R

Species Reactivity：

Human,Mouse,Rat,(predicted: Dog,Cow,Horse,)

Immunogen：

KLH conjugated synthetic peptide derived from human Legumain:201-300/433

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500Flow-Cyt=3ug/Test（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Placenta (Mouse) Lysate at 40 ugPrimary: Anti-Legumain (SL3907R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46 kDObserved band size: 55 kDSample: Kidney (Mouse) Lysate at 40 ugPrimary: Anti- Legumain (SL3907R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46 kDObserved band size: 55 kDSample: Lane 1: Human JAR cell lysates Lane 2: Human MCF-7 cell lysates Primary: Anti-Legumain (SL3907R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46 kDaObserved band size: 55 kDaParaformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Legumain) Polyclonal Antibody, Unconjugated (SL3907R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control: HL60. Primary Antibody (green line): Rabbit Anti-Legumain antibody (SL3907R) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG .Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.Overlay histogram showing Mouse spleen cells stained with SL3907-PE (red line). The cells were fixed with 1% paraformaldehyde (10 min).The cells were then incubated with the antibody (SL3907R-PE, 2ug/1x106cells) for 30 min at 22-25℃. Isotype control antibody (black line) was rabbit IgG (2ug/1x106cells) used under the same conditions. Acquisition of events were used for analysis.

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Unit:

Price: $228.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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This gene encodes a cysteine protease that has a strict specificity for hydrolysis of asparaginyl bonds. This enzyme may be involved in the processing of bacterial peptides and endogenous proteins for MHC class II presentation in the lysosomal/endosomal systems. Enzyme activation is triggered by acidic pH and appears to be autocatalytic. Protein expression occurs after monocytes differentiate into dendritic cells. A fully mature, active enzyme is produced following lipopolysaccharide expression in mature dendritic cells. Overexpression of this gene may be associated with the majority of solid tumor types. This gene has a pseudogene on chromosome 13. Several alternatively spliced transcript variants have been described, but the biological validity of only two has been determined. These two variants encode the same isoform. [provided by RefSeq, Jul 2008].

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.

Subcellular Location:
Lysosome (By similarity).

Tissue Specificity:
Ubiquitous. Particularly abundant in kidney, heart and placenta.

Post-translational modifications:
Glycosylated.

Similarity:
Belongs to the peptidase C13 family.

SWISS:
Q99538

Gene ID:
5641

Database links:

Entrez Gene: 5641 Human

Entrez Gene: 19141 Mouse

Entrez Gene: 63865 Rat

Omim: 602620 Human

SwissProt: Q99538 Human

SwissProt: O89017 Mouse

SwissProt: Q9R0J8 Rat

Unigene: 726036 Human

Unigene: 17185 Mouse

Unigene: 206021 Rat

Picture

Sample: Kidney (Mouse) Lysate at 40 ug
Primary: Anti- Legumain (SL3907R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kD
Observed band size: 55 kD

Sample:
Lane 1: Human JAR cell lysates
Lane 2: Human MCF-7 cell lysates
Primary: Anti-Legumain (SL3907R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 55 kDa

Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Legumain) Polyclonal Antibody, Unconjugated (SL3907R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: HL60. Primary Antibody (green line): Rabbit Anti-Legumain antibody (SL3907R) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PE Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Overlay histogram showing Mouse spleen cells stained with SL3907-PE (red line). The cells were fixed with 1% paraformaldehyde (10 min).The cells were then incubated with the antibody (SL3907R-PE, 2ug/1x10⁶cells) for 30 min at 22-25℃. Isotype control antibody (black line) was rabbit IgG (2ug/1x10⁶cells) used under the same conditions. Acquisition of events were used for analysis.

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