Legumain Polyclonal Antibody

Formalin-fixed and paraffin embedded human colorectal cancer labeled with Anti Legumain Polyclonal Antibody, Unconjugated (bs-3907R) followed by conjugation to the secondary antibody and DAB staining

$Lane 1: rat brain lysates Lane 2: rat heart lysates probed with Anti Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 in 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 49kD. Observed band size: 49kD.$

Lane 1: rat brain lysates Lane 2: rat heart lysates probed with Anti Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 in 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 49kD. Observed band size: 49kD.

$Moues kidney lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37\u00b0C.$

Moues kidney lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37\u00b0C.

$Mouse placenta lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37\u00b0C.$

Mouse placenta lysates probed with Legumain Polyclonal Antibody, unconjugated (bs-3907R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37\u00b0C.

HL-60 cells were fixed with 4% PFA for 10min at room temperature, 0.2% PBST for 20 min at room temperature. 5% BSA blocking buffer was used for 30 min at room temperature. Cells were then stained with Legumain Polyclonal Antibody (bs-3907R) at 1:33 dilution for 30 min at room temperature and then washed twice with 2% BSA in PBS. Secondary Antibody (PE-conjugated) incubation was performed for 40 min at room temperature. Acquisitions of 20,000 events were performed. The graph shows cells stained with primary antibody (green), and isotype control (orange).

Mouse spleen cells(black) were fixed with 4% PFA for 10min at room temperature, permeabilized with PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Legumain Polyclonal Antibody, A488 Conjugated (bs-3907R-A488) at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green) and isotype control (orange).

$Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (Legumain) Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

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\u00b0C for 30min; Antibody incubation with (Legumain) Polyclonal Antibody, Unconjugated (bs-3907R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Lane 1: Human JAR cell lysates; Lane 2: Human MCF-7 cell lysates probed with Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Human JAR cell lysates; Lane 2: Human MCF-7 cell lysates probed with Legumain Polyclonal Antibody, Unconjugated (bs-3907R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Horse

Overview
Catalog #	bs-3907R
Product Name	Legumain Polyclonal Antibody
Applications	WB IHC-P IHC-F IF FLOW-CYT
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Horse
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human Legumain
Immunogen Range	201-300/433
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5641
Swiss Prot	Q99538
Subcellular location	Cytoplasm
Synonyms	AEP; LGMN1; PRSC1; Legumain; Asparaginyl endopeptidase; Protease, cysteine 1; LGMN
Background	Has a strict specificity for hydrolysis of asparaginyl bonds. Can also cleave aspartyl bonds slowly, especially under acidic conditions. Required for normal lysosomal protein degradation in renal proximal tubules. Required for normal degradation of internalized EGFR. Plays a role in the regulation of cell proliferation via its role in EGFR degradation (By similarity). May be involved in the processing of proteins for MHC class II antigen presentation in the lysosomal/endosomal system.
Application Dilution
WB	1:300-5000
IHC-P	ELISA1:500-1000
IHC-F	FCM1:20-100
IF	IHC-P1:200-400
Flow-Cyt