Legumain Rabbit Polyclonal Antibody

SKU: orb5765

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Description

Research Area

Cell Biology, Hormone Biosynthesis

Images & Validation

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Item 1 of 6

Tested Applications	FC, IF, IHC-Fr, IHC-P, WB
Dilution range	WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=3ug/Test
Reactivity	Human, Mouse
Predicted Reactivity	Bovine, Canine, Equine, Rat

Related Conjugates & Formulations

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Key Properties

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Antibody Type	Primary Antibody
Host	Rabbit
Clonality	Polyclonal
Isotype	IgG
Immunogen	KLH conjugated synthetic peptide derived from human Legumain (201-300/433aa)
Target	LGMN
Molecular Weight	46 kDa
Purification	Affinity purified by Protein A
Conjugation	Unconjugated

Storage & Handling

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Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Form/Appearance	Liquid
Buffer/Preservatives	0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
Concentration	1mg/ml
Disclaimer	For research use only

Alternative Names

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

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Blank control: HL60. Primary Antibody (green line): Rabbit Anti-Legumain antibody (orb5765), dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min 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 20000 events was performed.

Overlay histogram showing Mouse spleen cells (red line). The cells were fixed with 1% paraformaldehyde (10 min). The cells were then incubated with the antibody (2 ug/1x106 cells) for 30 min at 22-25°C. Isotype control antibody (black line) was rabbit IgG (2 ug/1x106 cells) used under the same conditions. Acquisition of events were used for analysis.

Paraformaldehyde-fixed, paraffin embedded (mouse placenta), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Legumain) Polyclonal Antibody, Unconjugated (orb5765) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Kidney (Mouse) Lysate at 40 ug, Primary: Anti-Legumain (orb5765) 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 (orb5765) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46 kDa, Observed band size: 55 kDa.

Sample: Placenta (Mouse) Lysate at 40 ug, Primary: Anti-Legumain (orb5765) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46 kD, Observed band size: 55 kD.

Quick Database Links

Gene Symbol

LGMN

UniProt

Q99538

UniProt Details

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No UniProt data available

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Protocol Information

Western Blot (IB, immunoblot)

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IHC-P

Immunohistochemistry Paraffin

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IHC-Fr

Immunohistochemistry Frozen

Flow Cytometry

Immunofluorescence