Ab13970

Manufactured by R&D Systems

Ab13970 is an antibody product manufactured by R&D Systems. It is a polyclonal antibody raised against a specific antigen. The antibody can be used for research purposes, but its core function is not provided in this unbiased and factual description.

Automatically generated - may contain errors

Lab products found in correlation

Af566, by R&D Systems (2 mentions) Fluorescence tagged secondary antibodies, by Jackson ImmunoResearch (2 mentions) Af1835, by Merck Group (2 mentions) Sc 22458, by R&D Systems (2 mentions) Af2414, by Santa Cruz Biotechnology (2 mentions) Sc 206, by Abcam (2 mentions) Af5375, by R&D Systems (2 mentions) F7425, by Merck Group (2 mentions) Rabbit anti collagen 4, by Bio-Rad (1 mentions) Goat anti ve cadherin, by Santa Cruz Biotechnology (1 mentions) Mouse anti α smooth muscle actin cy3, by Merck Group (1 mentions) Rat anti endomucin v 7c7, by Santa Cruz Biotechnology (1 mentions) Af647, by Jackson ImmunoResearch (1 mentions) Af594, by Jackson ImmunoResearch (1 mentions) Rat anti pecam1, by BD (1 mentions) Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions) Donkey serum, by Merck Group (1 mentions) D9542, by Merck Group (1 mentions) Goat anti sox2, by R&D Systems (1 mentions) Alexa fluor 488 affinipure donkey anti chicken, by Jackson ImmunoResearch (1 mentions)

7 protocols using ab13970

Immunofluorescence Characterization of Taste Receptors

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Animals were euthanized and fixed by intracardiac perfusion with a 4% paraformaldehyde solution. Tongues were excised and placed in 30% sucrose solution overnight at 4°C for cryoprotection. Tissues were embedded in OCT compound and sectioned at 30 μm thickness on a cryostat. Sections were washed in PBS with 0.1% Triton X-100 (PBST), blocked with 10% donkey serum in PBST, incubated with primary antibodies overnight at 4°C, and incubated with fluorescence-tagged secondary antibodies (Jackson ImmunoResearch) for 2 hours at room temperature. Primary antibodies used were: anti-Sema7A (R&D Systems #AF1835; 1:300 dilution), anti-FLAG (Sigma #F7425; 1:1000 dilution), anti-T1R3 (Santa Cruz Biotechnology #sc-22458; 1:500 dilution), anti-Car4 (R&D Systems #AF2414; 1:500 dilution), anti-Plcβ2 (Santa Cruz Biotechnology #sc-206; 1:1000 dilution), anti-GFP (Abcam #ab13970; 1:500 dilution), anti-Nrp1 (R&D Systems #AF566; 1:200 dilution), and anti-PlxnC1 (R&D Systems #AF5375; 1:200 dilution).

Lee H., Macpherson L.J., Parada C.A., Zuker C.S, & Ryba N.J. (2017). Rewiring the Taste System. Nature, 548(7667), 330-333.

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Immunostaining Protocol for Mesenteric Vascular Structures

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Mesenteries were fixed in 4% paraformaldehyde at room temperature for 2 h and stained as previously described (Stanczuk et al., 2015 (link)). The following primary antibodies were used: mouse anti-α-smooth muscle actin-Cy3 (Sigma-Aldrich, C6198, 1:250), rat anti-endomucin (V.7C7) (Santa Cruz, SC-65495, 1:200), rat anti-CD41-FITC (eBioscience, 11-0411-81, 1:50), rabbit anti-collagen IV (Bio-Rad, 2150-1470, 1:500), chicken anti-GFP (Abcam, ab13970, 1:200), goat anti-neuropilin 2 (R&D Systems, AF567, 1:200), hamster anti-PECAM1 (Millipore, MAB1398Z, 1:1000), rat anti-PECAM1 (BD Pharmingen, 553370, 1:1000), rat anti-PECAM1-AF594 (BioLegend, 102520, 1:100), rabbit anti-human PROX1 (Stanczuk et al., 2015 (link); 1:200), rat anti-TER-119 (eBioscience, 145921, 1:200), rat anti-TER-119-AF647 (BioLegend, 116218, 1:50), goat anti-VE-cadherin (Santa Cruz, SC-6458, 1:200) and anti-podoplanin (Developmental Studies Hybridoma Bank at the University of Iowa, clone 8.1.1, 1:800). Autofluorescence signal at 550-600 nm wavelength was used for visualization of RBCs. Secondary antibodies conjugated to AF488, AF594, AF647 or Cy3 were from Jackson ImmunoResearch, and all were used at 1:300.

Zhang Y., Daubel N., Stritt S, & Mäkinen T. (2018). Transient loss of venous integrity during developmental vascular remodeling leads to red blood cell extravasation and clearance by lymphatic vessels. Development (Cambridge, England), 145(3), dev156745.

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Immunofluorescence Staining Protocol for Tissue Sections

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For both perfused or fresh frozen and post-fixed samples, sections were directly blocked for 1 h with blocking buffer (0.2% Triton-X (Sigma-Aldrich, X100), 3% Donkey serum (Merck, S30) in Tris-buffered saline (TBS, 50 mM Tris–Cl, pH 7.4, 150 mM NaCl), and incubated with the indicated primary antibodies in blocking buffer at 4 °C overnight. The following primary antibodies and dilutions were used: Chicken-anti GFP (1:500, abcam, ab13970), goat anti-SOX2 (1:500, R&D, AF2018) and rabbit anti-Iba1 (1:1,000, Fujifilm Wako, 019–19,741).
Sections were washed 3 times in TBS for 10 min and incubated with secondary antibodies at a concentration of 1:250 (Alexa Fluor 488 AffiniPure Donkey Anti-Chicken, JacksonImmuno Research, 703–545-155, and Alexa Fluor 594 AffiniPure Donkey Anti-Goat, JacksonImmuno Research 705-585-147) in blocking buffer at least 1 h at room temperature, protected from light. Slides were washed 2 times 10 min with TBS. Nuclei were stained with DAPI (1:5,000, Sigma-Aldrich, D9542) for 5 min. After other 3 washes of 10 min each, slides were mounted using self-made polyvinyl alcohol (PVA, Sigma P8136) mounting medium with 1, 4-diazabicyclo[2,2,2]octane (DABCO, Sigma D27802).

Scandella V., Paolicelli R.C, & Knobloch M. (2020). A novel protocol to detect green fluorescent protein in unfixed, snap-frozen tissue. Scientific Reports, 10, 14642.

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Immunofluorescence Characterization of Taste Receptors

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Lee H., Macpherson L.J., Parada C.A., Zuker C.S, & Ryba N.J. (2017). Rewiring the Taste System. Nature, 548(7667), 330-333.

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Barrel Cortex Development in Transgenic Mice

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VIPCre x Prox1eGFP (Control and cKO) animals age P4 and P6 were deeply anesthetized before transcardial perfusion with first 1× PBS followed by ice-cold 4% PFA. The brains were dissected and postfixed in ice-cold 4% PFA for 1 h, then cryo-protected in a 30% sucrose solution at 4°C for >24 h. The brains were embedded in OCT using a peel-away mold and stored at −80°C. Coronal 20-μm-think brain sections containing barrel cortex were cut and collected on-slide using a cryostat (Microm International, HM 560 M), and the slides were stored at −80°C until further processing.
The slides were thawed and washed using 1× PBS (3× 5min) before being blocked using 1.5% normal donkey serum (NDS) and 0.05% Triton X-100 in 1× PBS for 1 h. The primary antibodies GFP (chicken anti GFP, Abcam, ab13970) and Prox1 (goat anti Prox1, R&D Systems, AF2727) were both diluted 1:1000 in the blocking solution and left to incubate overnight at 4°C. The next day, the slides were washed (3× 7min) with 1× PBS before applying the secondary antibodies (donkey anti-goat 650, Thermo Fisher Scientific, SA5-10089 and donkey anti-chicken, Jackson ImmunoResearch Laboratories, 703-545-155) diluted at 1:1000 in 1× PBS for 2 h. The slides were coverslipped with Fluoromount-G with DAPI (00-4959-52) and imaged using an inverted Leica Microsystems SP8 microscope.

Stachniak T.J., Kastli R., Hanley O., Argunsah A.Ö., van der Valk E.G., Kanatouris G, & Karayannis T. (2021). Postmitotic Prox1 Expression Controls the Final Specification of Cortical VIP Interneuron Subtypes. The Journal of Neuroscience, 41(39), 8150-8162.

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Immunohistochemical Staining of Ocular Tissues

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Globes were enucleated and fixed by emersion (2% formaldehyde in phosphate buffered saline pH 7.5, overnight at 4°C). Fixed globes were then opened with a razor blade and processed through a sucrose gradient prior to embedding in OCT media (cryosections) or dehydrated and embedded in paraffin using an automated tissue processor (Leica). 7 μm cryosections or 5μm paraffin sections were then used for immunostaining using standard methods. Paraffin sections were dewaxed and subjected to heat induced antigen retrieval (10 mM TRIS, 1 mM EDTA, 0.05% Tween20, pH 9, autoclaved 121°C 30 min liquid cycle) prior to staining. For staining, all sections were first blocked and permeabilized (5% donkey serum, 2.5% BSA, 0.5% Triton X100 in TBS, pH 7.5, 1 hr at room temperature) and then incubated overnight with appropriate primary antibodies diluted in additional blocking buffer. Slides were then washed (6 × 5 minutes, 0.05% Tween 20 in TBS, pH 7.5) and incubated for an additional 1hr at room temperature with appropriate Alexafluor-conjugated secondary antibodies (Invitrogen, Carlsbad CA, USA). Primary antibodies used: Goat anti PODXL, 1:250 (R&D systems AF1556), Goat anti CD31, 1:250 (R&D systems AF3628), Chicken anti GFP 1:5000 (Abcam, AB13970), Goat anti PDGFRB 1:100 (R&D systems AF1042). Slides were mounted and imaged using a Nikon A1R confocal microscope.

Liu P., Lavine J.A., Fawzi A., Quaggin S.E, & Thomson B.R. (2022). Angiopoietin-1 is required for vortex vein and choriocapillaris development in mice. Arteriosclerosis, thrombosis, and vascular biology, 42(11), 1413-1427.

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Immunohistochemistry of GFP and CaMKIIa

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Mice were euthanized and transcardially perfused with PBS, immediately followed by perfusion with 4% paraformaldehyde (PFA). Brains were post-fixed overnight in PFA at 4°C and coronally sectioned with a cryostat (Leica) at 40 μm. Brain cryosections were incubated for 16 h at 4°C with primary antibody: anti-GFP (1:1000; Abcam, ab13970) and anti-CaMKIIa (1:500; R&D systems, MAB5584) antibodies in 10 % normal goat serum (NGS) in PBST (PBS, 1% BSA, and 0.5% Triton X-100). The sections were then washed and incubated for 2 hours at room temperature with the secondary Alexa 488 goat anti-chicken (1:400; Invitrogen, A11039) and 568 goat anti-mouse (1:400, Invitrogen, A11031) antibodies in 1% NGS/PBST. Stained sections were washed and DAPI-stained (Invitrogen). All sections were mounted and imaged on either Zeiss AX10 Observer D1 or a Zeiss LSM800 confocal microscope at Johns Hopkins University Microscope Facility.

Namkung H., Yukitake H., Fukudome D., Lee B.J., Tian M., Ursini G., Saito A., Lam S., Kannan S., Srivastava R., Niwa M., Sharma K., Zandi P., Jaaro-Peled H., Ishizuka K., Chatterjee N., Huganir R.L, & Sawa A. (2022). The miR-124-AMPAR pathway connects polygenic risks with behavioral changes shared between schizophrenia and bipolar disorder. Neuron, 111(2), 220-235.e9.

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