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Af3244

Manufactured by R&D Systems
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AF3244 is a lab equipment product offered by R&D Systems. It is a device designed for laboratory use, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach. Further information about the intended use or specific capabilities of this product is not available.

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Lab products found in correlation

Ab14917, by Abcam (1 mentions) Ab45688, by Abcam (1 mentions) Ab129051, by Abcam (1 mentions) Anti pdgfrβ, by BioLegend (1 mentions) Alexa fluor 594 conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Ab34710, by Abcam (1 mentions) Alexa fluor 488 conjugated anti goat igg, by Jackson ImmunoResearch (1 mentions) Evos fl auto 2 imaging system, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Anti zo 1, by Thermo Fisher Scientific (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Ab39260, by Abcam (1 mentions) Odyssey fc unit, by LI COR (1 mentions)

Close spelling variants of this product

Anti-podoplanin goat anti-mouse polyclonal antibody (AF3244,

4 protocols using af3244

1

Multiplex Immunofluorescence Profiling of Lymph Nodes

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Fresh LNs were embedded in tissue-freezing medium. Cryostat sections (8 μm thick) were cut for imaging by fluorescence confocal microscopy. The following primary Abs were used for tissue staining: anti-HEV MECA79 (sc-19602, SCBT), anti-Lyve-1 (ab14917, Abcam), anti-PDPN (AF3244, R&D Systems), anti-ER-TR7 (sc-73355, SCBT), anti-CD11b (101202, BioLegend), anti-PDGFRβ (136005, Biolegend), anti-NG2 (ab129051, Abcam), anti-RANKL (510002 Biolegend), anti-CD35 (NBP2-52667, Novus Biologicals), anti-MAdCAM (16-5997-85,eBioscience), anti-ZO-1 (61-7300, Invitrogen), anti-Collagen I (ab34710, Abcam), anti-Fibronectin (ab45688, Abcam), anti-LepR (L9536, Sigma-Aldrich). The following secondary Abs were used: Alexa Fluor 488-conjugated anti-rabbit IgG, Alexa Fluor 594-conjugated anti-rabbit IgG, Alexa Fluor 488-conjugated anti-rat IgG, Alexa Fluor 594-conjugated anti-rat IgG, Alexa Fluor 488-conjugated anti-goat IgG, and Alexa Fluor 594-conjugated anti-goat IgG (Jackson ImmunoResearch). The stained tissue sections were imaged using EVOS FL Auto 2 Imaging System (Thermo Fisher Scientific). For the quantification of images, all images were automatically processed using ImageJ (NIH) and split into RGB channels. Auto threshold was used to convert intensity values of the immunofluorescent stain into numeric data. DAPI (VECTASHIELD, Vector Laboratories) was used to stain the cell nuclei.
Jiang L., Yilmaz M., Uehara M., Cavazzoni C.B., Kasinath V., Zhao J., Naini S.M., Li X., Banouni N., Fiorina P., Shin S.R., Tullius S.G., Bromberg J.S., Sage P.T, & Abdi R. (2022). Characterization of Leptin Receptor+ Stromal Cells in Lymph Node. Frontiers in Immunology, 12, 730438.
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2

Quantifying Fibroblast Contraction and Elongation

Cited 1 time
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Control or PDPN KD FRCs were seeded at 10,000 per well in 150 μl collagen/matrigel matrix8 ,26 (link),28 (link). Gels were set at 37°C for 30 minutes then covered with cell culture medium. In some wells, the following were added to both the gel mix and medium: 10 μg/ml CLEC-2-Fc, 10 μM ROCK inhibitor (Y27632) or 10 μg/ml anti-PDPN antibody (R&D, AF3244). Contraction of the gel at day 3 was quantified as the ratio of contracted gel/original area and plotted relative to control. Gels were stained with TRITC-labelled phalloidin and DAPI for maximum length analysis (Imaris). The furthest points of each individual cell were measured in x,y,z coordinates and vectors calculated for comparison (Extended data figure 5).
Acton S.E., Farrugia A.J., Astarita J.L., Mourão-Sá D., Jenkins R.P., Nye E., Hooper S., van Blijswijk J., Rogers N.C., Snelgrove K.J., Rosewell I., Moita L.F., Stamp G., Turley S.J., Sahai E, & Sousa C.R. (2014). Dendritic Cells Control Fibroblastic Reticular Network Tension and Lymph Node Expansion. Nature, 514(7523), 498-502.
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3

Quantifying Fibroblast Contraction and Elongation

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Control or PDPN KD FRCs were seeded at 10,000 per well in 150 μl collagen/matrigel matrix8 ,26 (link),28 (link). Gels were set at 37°C for 30 minutes then covered with cell culture medium. In some wells, the following were added to both the gel mix and medium: 10 μg/ml CLEC-2-Fc, 10 μM ROCK inhibitor (Y27632) or 10 μg/ml anti-PDPN antibody (R&D, AF3244). Contraction of the gel at day 3 was quantified as the ratio of contracted gel/original area and plotted relative to control. Gels were stained with TRITC-labelled phalloidin and DAPI for maximum length analysis (Imaris). The furthest points of each individual cell were measured in x,y,z coordinates and vectors calculated for comparison (Extended data figure 5).
Acton S.E., Farrugia A.J., Astarita J.L., Mourão-Sá D., Jenkins R.P., Nye E., Hooper S., van Blijswijk J., Rogers N.C., Snelgrove K.J., Rosewell I., Moita L.F., Stamp G., Turley S.J., Sahai E, & Sousa C.R. (2014). Dendritic Cells Control Fibroblastic Reticular Network Tension and Lymph Node Expansion. Nature, 514(7523), 498-502.
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4

Western Blot Analysis of Protein Expression

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Western blotting was done as previously described (61 (link)). Chemiluminescence was detected in an Odyssey Fc unit (Licor). The following antibodies and dilutions were used: HRP-conjugated anti–β-actin antibody (MilliporeSigma, A3854HRP, 1:10000), anti-TRPV4 (rabbit, Abcam, ab 39260, 1:1000), anti–AQP-5 (rabbit, Alomone AQP-005, 1:1000), anti-Podoplanin (goat, R&D Systems, AF3244, 1:500), secondary anti-goat IgG (whole molecule) peroxidase (MilliporeSigma, A5420-1ML, 1:10000), and secondary anti-rabbit IgG peroxidase (POX) antibody (MilliporeSigma, A6154, 1:10000). One representative of three Western blots is shown in the figures.
Weber J., Rajan S., Schremmer C., Chao Y.K., Krasteva-Christ G., Kannler M., Yildirim A.Ö., Brosien M., Schredelseker J., Weissmann N., Grimm C., Gudermann T, & Dietrich A. (2020). TRPV4 channels are essential for alveolar epithelial barrier function as protection from lung edema. JCI Insight, 5(20), e134464.
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