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Mab5272

Manufactured by Merck Group
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MAB5272 is a monoclonal antibody product developed by Merck Group. It is a laboratory reagent intended for research use. The core function of MAB5272 is to serve as a detection tool for target analytes in various in vitro research applications.

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Lab products found in correlation

Af1166, by R&D Systems (3 mentions) Axio examiner lsm 780, by Zeiss (2 mentions) Alexa fluor 488 or 568, by Thermo Fisher Scientific (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Ab183024, by Abcam (1 mentions) Af566, by R&D Systems (1 mentions) Af4439, by R&D Systems (1 mentions) N5389, by Merck Group (1 mentions) Goat anti mouse alexa 568, by Thermo Fisher Scientific (1 mentions) Donkey anti rabbit hrp, by Jackson ImmunoResearch (1 mentions) Mab377, by Merck Group (1 mentions) Donkey anti goat hrp, by Jackson ImmunoResearch (1 mentions) Donkey anti rabbit alexa488, by Jackson ImmunoResearch (1 mentions) Goat anti rat alexa 568, by Thermo Fisher Scientific (1 mentions) Alexa488 goat anti chicken, by Thermo Fisher Scientific (1 mentions) Gfp 1020, by Agilent Technologies (1 mentions) Donkey anti rabbit alexa 568, by Thermo Fisher Scientific (1 mentions) Donkey anti mouse hrp, by Jackson ImmunoResearch (1 mentions) A21207, by Thermo Fisher Scientific (1 mentions) Ab9260, by Merck Group (1 mentions)

5 protocols using mab5272

1

Antibody Characterization for Neuronal Proteins

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Primary antibodies used for immunohistochemistory and western blot were as follows; goat anti-NOVA2 (C-16) (sc-10546, Santa Cruz), rabbit anti-NOVA1 [EPR13847] (ab183024, abcam), human anti-pan NOVA (anti-Nova paraneoplastic human serum), rabbit anti-PTBP2 (Polydorides et al., 2000 (link)), rat anti-L1 (MAB5272, Millipore), goat anti-TAG1/Contactin-2 (AF4439, R&D systems), goat anti-NetrinG1a (AF1166, R&D systems), rabbit anti-NURR1 (M-196) (sc-5568, Santa Cruz), rabbit anti-neurofilament (AB1981, Chemicon), mouse anti-NF200 (N5389, SIGMA), mouse anti-ctbp2 (612044, BD), rabbit anti-myoVI (sc-50461, Santa Cruz), and goat anti-neuropilin-1 (AF566, R&D systems). Anti-NOVA1 and anit-NOVA2 antibody specificity for immunohistochemistory and immunoprecipitation was confirmed (Figure 1—figure supplement 2).
Saito Y., Miranda-Rottmann S., Ruggiu M., Park C.Y., Fak J.J., Zhong R., Duncan J.S., Fabella B.A., Junge H.J., Chen Z., Araya R., Fritzsch B., Hudspeth A.J, & Darnell R.B. (2016). NOVA2-mediated RNA regulation is required for axonal pathfinding during development. eLife, 5, e14371.
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2

Immunofluorescence and Western Blotting Protocols

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Following primary antibodies were used for Immunofluorescence (IF) and Western blotting (WB): rabbit anti-GFP (1:200 IF, Invitrogen, A11122), chicken anti-GFP (1:200 IF, Aves, GFP-1020), rabbit anti-GFAP (1:100 IF, Dako, Z0334), mouse anti-NeuN (1:50 IF, Millipore, MAB377), rat anti-L1 (1:100 IF, Millipore, MAB5272), rabbit anti-Calretinin (1:500 IF, Millipore, MAB5054), goat anti-Nrp1 (2 µg/ml for function blocking, R and D, AF566), goat anti-VEGFR2 (1:15 IF, R and D, AF644), rabbit anti-VEGFR2 (1:1000 WB, Cell Signaling, 2479), rabbit anti-(phospho)VEGFR2 (Y1175) (1:500 WB, Cell Signaling, 2478), rabbit anti-(phospho/SFK (Y416) (1:200 IF, Invitrogen, 44660G), mouse anti-beta-III-tubulin (1:150 IF, Sigma, T5076), goat anti-VE-Cadherin (1:1000 WB, R and D, AF1002), Phalloidin (1:400 IF, Sigma, P1951). The following secondary antibodies were used: donkey anti-rabbit Alexa488 (Jackson Immunoresearch, 711-545-152), donkey anti-rabbit Alexa568 (Life Technologies, A10042), donkey anti-goat Alexa568 (Molecular Probes, A11057), goat anti-mouse Alexa568 (Invitrogen, A11031), goat anti-rat Alexa568 (Invitrogen, A11077), goat anti-chicken Alexa488 (Molecular Probes, A11039), donkey anti-goat HRP (Jackson Immunoresearch, 705–0350147), donkey anti-rabbit HRP (Jackson Immunoresearch, 711-035-152), donkey anti-mouse HRP (Jackson Immunoresearch, 715-035-150).
Luck R., Urban S., Karakatsani A., Harde E., Sambandan S., Nicholson L., Haverkamp S., Mann R., Martin-Villalba A., Schuman E.M., Acker-Palmer A, & Ruiz de Almodóvar C. (2019). VEGF/VEGFR2 signaling regulates hippocampal axon branching during development. eLife, 8, e49818.
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3

Immunohistochemical Analysis of Netrin-G1a and L1 in E14.5 Mouse Brain

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E14.5 embryos were fixed in 4% paraformaldehyde for 24 hrs at 4°C, cryoprotected in 30% sucrose 24 hrs at 4°C and sectioned at 10 μm using a Leica CM1950 cryostat. Sections were blocked with 10% donkey serum for 1 hr. Primary antibodies were diluted in 3% donkey serum and incubated for 15–18 hrs at 4°C with Netrin-G1a anti-goat antibody (1:100, R&D Systems, AF1166) or Neural Cell Adhesion Molecule L1 anti-rat antibody (1:500, EMD Millipore, MAB5272). Sections were then incubated for 2 hrs at room temperature in their corresponding donkey anti-goat and anti-rat secondary antibodies conjugated to Alexa Fluor 568 or 488 (1:1000, Thermofisher Scientific). Images were acquired using the Zeiss Axio Examiner LSM 780 confocal microscope with 405nm (2%), 488nm (5%), and 561nm (8%) lasers. Rostral to caudal embryonic brain sections were scanned using Zen Blue and Black Edition 2012 software, 20X objective at 1.5X zoom with 5 × 1μm interval z-slices and 3 individual tracks for each fluorescent dye. Image acquisition settings included: scan mode set at frame, frame size set at 1024 X 1024, scan speed set at 7, averaging at 2 by line and mean, and bit depth set at 8 bit. Pinhole was set to 1AU.
Vuong H.E., Pronovost G.N., Williams D.W., Coley E.J., Siegler E.L., Qiu A., Kazantsev M., Wilson C.J., Rendon T, & Hsiao E.Y. (2020). The maternal microbiome modulates fetal neurodevelopment in mice. Nature, 586(7828), 281-286.
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4

Immunohistochemical Analysis of Netrin-G1a and L1 in E14.5 Mouse Brain

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E14.5 embryos were fixed in 4% paraformaldehyde for 24 hrs at 4°C, cryoprotected in 30% sucrose 24 hrs at 4°C and sectioned at 10 μm using a Leica CM1950 cryostat. Sections were blocked with 10% donkey serum for 1 hr. Primary antibodies were diluted in 3% donkey serum and incubated for 15–18 hrs at 4°C with Netrin-G1a anti-goat antibody (1:100, R&D Systems, AF1166) or Neural Cell Adhesion Molecule L1 anti-rat antibody (1:500, EMD Millipore, MAB5272). Sections were then incubated for 2 hrs at room temperature in their corresponding donkey anti-goat and anti-rat secondary antibodies conjugated to Alexa Fluor 568 or 488 (1:1000, Thermofisher Scientific). Images were acquired using the Zeiss Axio Examiner LSM 780 confocal microscope with 405nm (2%), 488nm (5%), and 561nm (8%) lasers. Rostral to caudal embryonic brain sections were scanned using Zen Blue and Black Edition 2012 software, 20X objective at 1.5X zoom with 5 × 1μm interval z-slices and 3 individual tracks for each fluorescent dye. Image acquisition settings included: scan mode set at frame, frame size set at 1024 X 1024, scan speed set at 7, averaging at 2 by line and mean, and bit depth set at 8 bit. Pinhole was set to 1AU.
Vuong H.E., Pronovost G.N., Williams D.W., Coley E.J., Siegler E.L., Qiu A., Kazantsev M., Wilson C.J., Rendon T, & Hsiao E.Y. (2020). The maternal microbiome modulates fetal neurodevelopment in mice. Nature, 586(7828), 281-286.
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5

Immunofluorescence Staining of Frozen Tissue

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Frozen sections were washed in PBS with 0.2% Triton X-100 (PBST) and blocked with 5% normal donkey serum (NDS) for 1 h. The slides were then incubated with primary antibodies mixed in 1% NDS overnight at 4°C. Antibodies against TCF7L2 (1:500; 2569, Cell Signaling Technology), Cav3.1 (1:500; MABN464, Sigma-Aldrich, NeuroMab clone N178A/9), β-galactosidase (1:100; AB986, Merck Millipore), L1CAM (1:500; MAB5272, Merck Millipore), PAX6 (1:100; PRB-278P, BioLegend), KI-67 (1:100; AB9260, Merck Millipore), TUJ1 (1:65; MAB1637, Merck Millipore), NKX2-2 (1:50; 74.5A5, Developmental Studies Hybridoma Bank), SIX3 (1:100; 200-201-A26S, Rockland), POU4F1 (1:300; Fedtsova and Turner, 1995 (link)) were used. Sections were then incubated for 1 h with appropriate secondary antibody conjugated with Alexa Fluor 488 or 594 (1:500; A-21202, A-21207 and A-11076, Thermo Fisher Scientific). The slides were additionally stained with Hoechst 33342 (1:10,000; 62249, Thermo Fisher Scientific), washed and mounted with Vectashield Antifade Mounting Medium (H1000, Vector Laboratories).
Lipiec M.A., Bem J., Koziński K., Chakraborty C., Urban-Ciećko J., Zajkowski T., Dąbrowski M., Szewczyk Ł.M., Toval A., Ferran J.L., Nagalski A, & Wiśniewska M.B. (2020). TCF7L2 regulates postmitotic differentiation programmes and excitability patterns in the thalamus. Development (Cambridge, England), 147(16), dev190181.
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