13-4400

antibody from Invitrogen Antibodies

Targeting: HSPG2 perlecan, PRCAN, SJS1

Provider product page for 13-4400

Western blot

ELISA

Immunohistochemistry

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Antibody data

Antibody Data
Antigen structure
References [19]
Comments [0]
Validations
Other assay [10]

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Product number: 13-4400 - Provider product page
Provider: Invitrogen Antibodies
Product name: Perlecan Monoclonal Antibody (7B5)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 7B5
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

HSPG2 protein structure - 13-4400 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 10 Double immunofluorescence staining for perlecan, 3-nitrotyrosine epitopes and CD14-positive cells in human atherosclerotic lesion sections. Frozen sections (5 mum) of human atherosclerotic lesions were incubated with monoclonal or polyclonal primary antibodies: anti-nitrotyrosine (rabbit IgG), anti-human perlecan (mouse mAb, clone 7B5, perlecan domain III) anti-human CD14 (mouse mAb), non-immune rabbit IgG and non-immune mouse IgG, and subsequently the following detection antibodies: goat anti-rabbit cyanine-3 (Cy-3)-labelled IgG or goat anti-mouse Cy-2-labeled IgG. DAPI was used to image cell nuclei. Images were acquired as described in the Materials and Methods. A) Heavily thickened intima of an artery with a type III/IV lesion. Epitopes for 3-nitrotyrosine (red signal) show marked co-localization with perlecan (green) present in the basement membrane of the endothelium (arrow) as well as with those of the vasa vasorum (dotted arrow). B) The signal for 3-nitrotyrosine (red) was frequently co-localized with CD14-positive cells (macrophages, green). The red signal for nitrotyrosine underneath the bulk of macrophages results from multiple oblique sections through a vas vasorum. C) In sections where the antibodies for perlecan/CD14 and for 3-nitrotyrosine were replaced with non-immune mouse IgG (control, green) and non-immune rabbit IgG (control, red), no staining in the intima was observed. The faint background staining in the media (also observed when no primary or se

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 Structural consequences of perlecan modification by peroxynitrite. Perlecan (330 nM) in 0.1 M phosphate buffer, pH 7 (lane 1) was exposed for 20 min at 22 degC to peroxynitrite at molar ratios of 250 (lane 2), 500 (lane 3), 1000 (lane 4), 1000 in the presence of 25 mM bicarbonate (lane 5), heparinase III for 16 h at 37 degC (lane 6), heparinase III followed by 1000-fold molar excess of peroxynitrite (lane 7), and dONOO (lane 8), prior to separation on 3-8% Tris-acetate gels under reducing conditions for 1 h at 150 V and subsequent western blotting to nitrocellulose. (A) Stains-all/silver stain of gel. (B) Western blot probed for 3-nitroTyr formation with mAb against 3-nitroTyr (HM11). (C) Western blot probed for heparan sulfate epitopes with mAb 10E4. (D) Western blot probed for perlecan domain III with mAb 7B5. Position of molecular mass markers are shown for reference: < perlecan heparan sulfate, < perlecan protein core.

Supportive validation

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Experimental details: Figure 1 Anion Exchange Chromatography of rhPln.247 conditioned media (CM) . A: Chromatogram of elution profile for rhPln.198-Ad and rhPln.247-Ad CM expressed from HEK 293 cells; B and C: Western blots of anion exchange fractions with mAb CSI 001-71 recognizing the Pln.D1 core; D and E: Stains-All analysis of HEK 293 fractions; F: Fractions from HUVEC synthesis. Panels B and D represent rhPln.198 fractionation; panels C, E, F represent rhPln.247 fractionation. (M) Markers; (CM) Conditioned medium undiluted from cells. Similar data were generated using plasmid expression of pln.247 (not shown).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 ELISA identifies only Pln.D1 but not Pln.D3-5 in enriched rhPln.198 . The enriched high M r rhPln.198 synthesized by HEK 293 cells was used to coat microtiter wells that were subsequently blocked then incubated with 1.7 mug/ml of different primary antibodies as depicted in the legend. ELISA was completed as described in Methods. Background signal for each antibody binding to uncoated wells was subtracted to produce the net signal. Inset: ELISA data showing negligible mAb A76 reactivity against rhPln.247 and strong reactivity with full-length endothelial perlecan (purified as previously described [ 53 ]), while mAb A71 reacted strongly to both the native perlecan and the truncated recombinant. Net absorbance minus buffer-coated wells represented.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 Immunoblot analysis of growth factor binding to Pln.198 and Pln.247 . 500 ng growth factors were bound to nitrocellulose, the blots were blocked, and then incubated in either 1 ug/ml Pln.198 or Pln.247 (panel A) or rhPln.247 pre-digested with heparinase III (panel B). Detection was performed with anti-perlecan domain 1 primary antibody CSI 001-71. Note that while both the background BSA signal and the internal control rhPln.247 signals are slightly higher in panel B (bottom two samples), a decrease in the growth factor binding to Pln.247 was present after digestion.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Decellularization of human bronchial airways efficiently removes cells while preserving the extracellular matrix. ( A) The macroscopic structure of the 500 um thick human bronchial tissue sections was preserved after decellularization (DC). (B) Hematoxylin & eosin, Masson's trichrome (collagen in blue) and alcian-blue periodic acid Schiff (AB-PAS) (polysaccharides in magenta/purple) stainings as well as immunohistochemistry against the basement membrane proteins collagen IV, laminin and perlecan, before and after DC. Images are representative of n = 3. (C) Dye-binding methods confirmed that DC efficiently decreased DNA content (**p = 0.002) while preserving sulfated glycosaminoglycans (sGAG) and elastin in bronchial scaffolds (n = 6 for DNA, n = 3 for sGAG and elastin). The data were analyzed with the Mann-Whitney test and graphs indicate mean and standard deviation. Scale bars: 3 mm in A and 50 um in B.