- 53-9871-80 - Invitrogen Antibodies COL4A5 antibody

Submitted references SeqStain is an efficient method for multiplexed, spatialomic profiling of human and murine tissues.

Rajagopalan A, Venkatesh I, Aslam R, Kirchenbuechler D, Khanna S, Cimbaluk D, Kordower JH, Gupta V
Cell reports methods 2021 Jun 21;1(2)

Spontaneous Extracellular Matrix Accumulation in a Human in vitro Model of Renal Fibrosis Is Mediated by αV Integrins.

Bon H, Hales P, Lumb S, Holdsworth G, Johnson T, Qureshi O, Twomey BM
Nephron 2019;142(4):328-350

Six Weeks of Low-Load Blood Flow Restricted and High-Load Resistance Exercise Training Produce Similar Increases in Cumulative Myofibrillar Protein Synthesis and Ribosomal Biogenesis in Healthy Males.

Sieljacks P, Wang J, Groennebaek T, Rindom E, Jakobsgaard JE, Herskind J, Gravholt A, Møller AB, Musci RV, de Paoli FV, Hamilton KL, Miller BF, Vissing K
Frontiers in physiology 2019;10:649

A high content, phenotypic 'scar-in-a-jar' assay for rapid quantification of collagen fibrillogenesis using disease-derived pulmonary fibroblasts.

Good RB, Eley JD, Gower E, Butt G, Blanchard AD, Fisher AJ, Nanthakumar CB
BMC biomedical engineering 2019;1:14

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry of formalin-fixed paraffin embedded human placenta using 10 µg/mL Anti-Human Collagen Type IV Alexa Fluor® 488. Nuclei are stained with DAPI.
Conjugate: Green dye

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Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 Markers of myofibroblast activation and extracellular matrix deposition. a Immunocytochemical analysis of fibroblast activation (alpha-SMA) and ECM deposition (collagen type I, collagen type IV and fibronectin) in response to TGF-beta 1 stimulation [1 ng/ml] for 72 h. Images are representative of n = 3 independent experiments at 10x magnification. Scale bars represent 200 mum. b mean fluorescent intensity of markers in (A) expressed as percent mean fluorescent intensity (MFI) normalised to ficoll treated-vehicle control. Histogram represents mean +- SEM. ***P
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3. Enzymatic de-staining of tissues stained using the SeqStain antibodies Immunofluorescence images showing human kidney tissue sections stained using SeqStain antibodies (as indicated in the panel). The antibodies were labeled using either the AF488 fluorophore (shown in green), the Cy3 fluorophore (shown in red), or the Cy5 fluorophore (shown in yellow). Immunofluorescence images of these tissue sections after de-staining with DNase I treatment are shown below each panel. All images are representative of at least three replicates. Graphs showing quantification of fluorescence intensity after staining (red bars, green bars, or yellow bars) and de-staining (brown bars) in each panel is also presented on the right. Graphs show the mean +- SD. Scale bar, 100 mum.
Conjugate: Green dye

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Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4. Selective de-staining of complex tissues using SeqStain antibodies and restriction endonucleases (A) Schematic representation of the technique for selective removal of fluorophores from immunofluorescently labeled tissues. Tissue can be stained with a combination of SeqStain antibodies labeled using fluorescent DNA that contains specific recognition sites for restriction endonuclease(s) (such as EcoRV and SmaI). Treatment of samples with the specific restriction nuclease selectively removes fluorophores only from the antibodies carrying the respective DNA sequence, leaving all others undiminished. (B) Immunofluorescence images showing normal human kidney tissue sections after each of the cycles of staining with unique SeqStain antibodies (as indicated in the panel) and de-staining with a specific restriction endonuclease (as indicated). The antibodies were labeled using the AF488 fluorophore (shown in green) or the Cy3 fluorophore (shown in red). Merged images from the two fluorescence channels (along with images from DAPI-stained nuclear markers) are also shown. All images are representative of at least three replicates. Graph showing quantification of fluorescence intensity after staining (green and red bars) and de-staining (brown bars) in each panel is also presented on the right. Graphs show the mean +- SD. Scale bars, 100 mum.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. SeqStain-based multiplex imaging of whole human kidney tissue provides a 20-plex image Immunofluorescent images of whole kidney tissue sections after each round of staining with unique SeqStain antibodies (as indicated) and DAPI (as indicated in the panel). Zoomed-in sections of images are presented below each panel. A serial section stained with H&E is also presented. Scale bars, 100 mum.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7. SeqStain multiplex imaged panels identify major substructures in the human kidney (A) Image showing a composite overlay of aligned immunofluorescence image stack of 20 unique markers from the whole kidney tissue section. Scale bar, 100 mum. (B) Image of a zoomed-in region from the whole kidney tissue section in (A) (white square) showing various components of the kidney tissue, including the glomerular endothelial cells (GECs), proximal tubule (PT), collecting duct (CD), distal convoluted tubules (DCT), and podocytes. Scale bar, 100 mum. (C) Boxed dot plot showing computed co-expression of EpCAM and the indicated markers on a per-cell basis (mean +- SD). (D) Image of a zoomed-in region from the whole kidney tissue section in (A) (red square) showing one glomerulus (two panels), false colored for the indicated markers. Scale bar, 100 mum. (E) Representative images showing zoomed-in regions of composite overlay of aligned immunofluorescence image stacks from (A) for the identification of various immunophenotypes of cells and tissue sections based on co-localization of various markers (as labeled). Scale bar, 50 mum. (F) Boxed dot plot of cellular neighborhoods showing the computed distances of the indicated cells from glomerular basement membrane of a selected glomerulus in the kidney (mean +- SD). Glom refers to cells residing inside the glomerulus, whereas RI refers to cells residing outside the glomerulus, in the renal interstitium.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5. Both epithelial cells and fibroblasts contribute to the accumulation of ECM. a Images show immunofluorescent staining of the epithelial cells (Cell Trace TM Violet, blue) and fibroblasts (Cell Trace TM Yellow, orange) for fibronectin, collagens I, III, IV, V or VI (green) after 1 or 7 days in co-culture. b Epithelial cells and fibroblasts were cultured for 48 h in mono-culture or co-culture. RNAs were then extracted and transcript expression levels were assessed using Affymetrix Bead Arrays. The heat map shows the expression levels of a panel of ECM genes in the mono-cultures and the co-culture. Results are summarised in Table 1, signal value (-) 1,000. E, mono-culture of epithelial cells; F, mono-culture of fibroblasts; E + F, co-culture of epithelial cells and fibroblasts. Images show a single field. Scale bars represent 50 um. ECM, extracellular matrix.
Conjugate: Green dye

53-9871-80

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