PA1-1746

antibody from Invitrogen Antibodies

Targeting: ACAN AGC1, CSPG1, CSPGCP, MSK16

Western blot

Immunohistochemistry

Antibody data

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Validation data

Product number: PA1-1746 - Provider product page
Provider: Invitrogen Antibodies
Product name: Aggrecan Neo Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-1746 detects human, rat, mouse, bovine, canine, and porcine aggrecan neo G1. PA1-1746 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~64 kDa protein representing the neoepitope sequence (NITEGE) generated by aggrecanase-mediated cleavage at Glu373-Ala374 of aggrecan core proteins. The PA1-1746 immunogen is a synthetic peptide corresponding to residues residues C G G N(387) I T E G E(392) of human aggrecan.
Reactivity: Human, Mouse, Rat, Bovine, Canine, Porcine
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.83 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

ACAN protein structure - PA1-1746 shown in red.

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 HT/PCy supplementation reduces the severity of mid-stage OA in ACLT rabbits. Rabbits (n = 6 per group) underwent ACLT of the right knee. NaCl (Std), GlcN or HT/PCy were orally administrated every two days for 13 weeks starting 3 weeks before surgery. HES staining of cartilage ( A ) and OARSI scores ( B ) of non-operated knee (control) or operated knee (ACLT) at 10 weeks post-surgery are shown. Immunohistochemical detection of aggrecan epitopes (NITEGE) ( C ) and relative staining intensity of the articular cartilage matrix ( D ) of standard diet (Std), GlcN and HT/PCy diet groups at 10 weeks after ACLT. (i) Non calcified-cartilage, (ii) calcified cartilage and (iii) sub-chondral bone. *p > 0.05 compared to non-operated knee (control) within the same group; # p > 0.05 compared to ACLT receiving NaCl (Std) group, $ p > 0.05 compared to ACLT receiving GlcN. Bar represents 250 mum ( A ) and 100 mum ( C ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. ADAMTS proteoglycanases are highly expressed and active in the human umbilical vein. ( a ) RNA in situ hybridization shows robust ADAMTS1 and ADAMTS9 expression in umbilical vein endothelium and tunica media (TM) and in outer arterial TM. Robust ADAMTS4 and ADAMTS5 expression was confined to the venous endothelium, with moderate ADAMTS4 expression and minimal ADAMTS5 expression in SMC (n = 3 umbilical cords for each probe). ( b ) ADAMTS-cleaved aggrecan (anti-NITEGE, red) and versican (anti-DPEAAE, red) both showed strong ADAMTS proteolytic activity throughout the venous wall and the outer artery TM. Unlike aggrecan, extensive versican proteolysis is seen in the arterial intima and sub-intima (n = 4 umbilical cords for each antibody). Wj, Wharton''s jelly. The brackets mark TM boundaries. Scale bars in a-b = 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. Aggrecan enrichment in the inner umbilical artery tunica media (TM) and its proteolysis in the umbilical vein is a characteristic of large mammals. ( a ) Alcian blue-eosin staining of umbilical cord sections shows proteoglycan enrichment (blue) in the inner arterial tunica media (TM). The elephant umbilical vein was unavailable. ( b ) Anti-CS immunofluorescence (7D4, green) shows enrichment in the inner arterial TM. Bonobo cords lacked 7D4 reactivity. ( c,d ) Aggrecan and anti-NITEGE immunostaining from reactive species showed aggrecan enrichment in the inner arterial TM and aggrecan proteolysis in the vein and outer artery TM. n = 3 for Gazelle and n = 1 for other mammals. Triplicate sections were stained from each animal cord. Scale bars = 100 mum and 200 mum. * indicates the vessel lumen. Figure 3-figure supplement 1. Aggrecan cleavage site conservation in mammals. The cleavage site location is shown over the sequence alignment by scissors, and the critical Glu(E) residue essential for cleavage is shown by red highlighting. Bold text identifies the cleavage-revealed neo-epitope NITEGE. Residue numbers are indicated for human and mouse aggrecan only.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6. Reduced aggrecan and versican proteolysis in Adamts1 -/- umbilical vessels. ( a,b ) E17.5 Adamts1 -/- umbilical vessels show increased aggrecan staining and reduced anti-NITEGE staining in ( a ), quantified in ( b ) (n = 3 cords each genotype, error bars indicate mean +-S.D. *, p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Chondrogenic effects of FRZB on BMSCs in vitro . ( A ) Alcian blue staining of BMSCs in different treatment groups at 2 weeks to indicate GAG production in the culture plate. ( B ) Quantification of GAG production in generated cartilaginous tissues (n=6 for each). ( C ) Immunofluorescent assay of Aggrecan (green) expression and nucleus (blue) in different treatment groups observed under confocal microscopy. ( D - F ) Expression level of chondrogenic markers for BMSCs in different treatment groups (n=6 for each) in ( A ) (n=6 for each). *P < 0.05 between control group and other groups. Data are presented as averages +- SD. One-way analysis of variance (ANOVA) with post-hoc Tukey''s B test was applied. Ab, antibody; Exo, exogenous; ACAN , aggrecan; GAG, glycosaminoglycan.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6. Aggrecan in the human vasculature. A , Localization of aggrecan, the aggrecan NITEGE, and versican DPEAAE neoepitopes (neo), and HPLN1 in stented and control human coronary arteries with the presence of atherosclerosis (&ast). Arrows mark the contacts of the stent struts with coronary artery. Scale bars=1 mm. B , Colocalization of aggrecan (Alexa 633, displayed in green) and aggrecan NITEGE neoepitope (Alexa 568, displayed in red) in human stented coronary arteries by immunofluorescence. Note the presence of aggrecan fragments at the contacts of the stent struts with the artery and in the subendothelial layer (zoomed-in areas). Overview image 20x, scale bar=500 um; Zoomed-in areas 60x, scale bar=25 um. C , Adjusted peak area for aggrecan, versican, and decorin in the human thoracic aorta and saphenous veins as determined by targeted proteomics. n=7 per group. &ast P