Log In Sign up
The largest database of trusted experimental protocols

Mrb 435p

Manufactured by Fortrea
Visit Supplier
Sourced in United States

The MRB-435P is a laboratory equipment designed for general scientific applications. It serves as a multipurpose tool for various experimental and analytical tasks. The core function of this product is to provide a reliable and versatile platform for researchers and scientists to conduct their work effectively.

Automatically generated - may contain errors

Lab products found in correlation

Mab5326, by Merck Group (4 mentions) Mms 435p, by Merck Group (3 mentions) Axiovert 200m microscope, by Zeiss (2 mentions) Laminin, by Thermo Fisher Scientific (2 mentions) E13.5 cd 1 embryos, by Charles River Laboratories (2 mentions) 35 cm glass bottom dishes, by MatTek (2 mentions) Goat anti dlx3, by Santa Cruz Biotechnology (2 mentions) Ab74065, by Abcam (2 mentions) Mouse anti β actin, by Merck Group (2 mentions) Ab13970, by Fortrea (2 mentions) Dm5000b microscope, by Leica (2 mentions) Mab374, by Merck Group (2 mentions) Mab3402, by Merck Group (2 mentions) Anti map2 antibody, by Merck Group (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (2 mentions) Alexa fluor 594, by Thermo Fisher Scientific (2 mentions) Nestin, by Merck Group (2 mentions) Ab5622, by Merck Group (2 mentions) Mab1637, by Merck Group (2 mentions) Confocal imaging system, by Leica (2 mentions)

21 protocols using mrb 435p

1

Time-lapse Imaging of Neuronal Transdifferentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
MEFs were isolated from E13.5 CD-1 embryos (Charles River) and infected with a dox-inducible, N-terminal-tagged EGFP-Ascl1 fusion construct using the protocol previously described1 (link). Cells were plated on 35 cm glass bottom dishes (MatTek), coated with polyorthinine (Sigma P3655) and laminin (Invitrogen 23017-015). Imaging experiments were performed between 3 and 6 days post-dox induction, in a temperature and CO2-controlled chamber. Images were taken for up to 10 positions per dish, for 3 dishes, every 45 minutes with a Zeiss AxioVert 200M microscope with an automated stage using an EC Plan-Neofluar 5x/0.16NA Ph1 objective or an A-plan 10x/0.25NA Ph1 objective. Cells were fixed at 6 days and immunostained using Tuj1 antibodies recognizing the neuronal β-3-tubulin Tubb3 (Covance MRB-435P) to confirm neuronal identity. We used ImageJ to segment individual cells and measure the level of GFP for 7 Tuj1+ cells and 7 Tuj1− cells over time. Average intensity was obtained by normalizing the average intensity of a cell segment by the average background intensity of an adjacent segment of the same size. A t-test was performed comparing Tuj1+ and Tuj1− cells at each time-point to evaluate significance.
Treutlein B., Lee Q.Y., Camp J.G., Mall M., Koh W., Shariati S.A., Sim S., Neff N.F., Skotheim J.M., Wernig M, & Quake S.R. (2016). Dissecting direct reprogramming from fibroblast to neuron using single-cell RNA-seq. Nature, 534(7607), 391-395.
+ Open protocol
+ Expand
2

Immunofluorescence Staining of Neural Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rabbit anti-Ascl1 (Abcam ab74065), chicken anti-GFP (Abcam ab13970), rabbit anti-Tubb3 (Covance MRB-435P), mouse anti-Tubb3 (Covance MMS-435P), mouse anti-Map2 (Sigma M4403), rabbit anti-Myh3 (Santa Cruzsc-20641), goat anti-Dlx3 (Santa Cruz sc-18143), mouse anti-β-Actin (Sigma A5441), rabbit anti-Tcf12 (Bethyl A300-754A).
Treutlein B., Lee Q.Y., Camp J.G., Mall M., Koh W., Shariati S.A., Sim S., Neff N.F., Skotheim J.M., Wernig M, & Quake S.R. (2016). Dissecting direct reprogramming from fibroblast to neuron using single-cell RNA-seq. Nature, 534(7607), 391-395.
+ Open protocol
+ Expand
3

Immunoblotting and Immunostaining Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunoblotting, a polyclonal rabbit antibody (Calarco et al. 2009 (link)) raised against amino acids 1–82 of nSR100 was used at 1:5000. Anti-tubulin (T6074, Sigma) was used at 1:5000. For immunostaining, mouse monoclonal anti-neurofilament (2H3 conditioned medium, Iowa Developmental Studies Hybridoma Bank) was diluted to 1:50 for whole-mount diaphragm staining and 1:100 for brain section staining. Mouse anti-NeuN (mab377, Millipore), mouse anti-Satb2 (ab51502, Abcam), rabbit anti-Tbr1 (ab31940, Abcam), and chicken polyclonal anti-β-galactosidase (ab9361, Abcam) were all diluted to 1:500. Chicken anti-MAP2 (ab5392, Abcam) was diluted to 1:10,000, mouse anti-Tuj1 (MRB-435P, Covance) was diluted to 1:750, and rabbit anti-Pax6 (PRB-278P, Covance) was diluted to 1:1500. For in situ hybridization, an anti-DIG antibody conjugated to alkaline phosphatase (Roche) was diluted to 1:5000.
Quesnel-Vallières M., Irimia M., Cordes S.P, & Blencowe B.J. (2015). Essential roles for the splicing regulator nSR100/SRRM4 during nervous system development. Genes & Development, 29(7), 746-759.
+ Open protocol
+ Expand
4

Primary Neuron Culture and Progranulin Effects

Check if the same lab product or an alternative is used in the 5 most similar protocols
Preparation of primary cultures was performed as described58 (link). Briefly, cortices from P0 or P1 rat pups were isolated and plated at 160,000 cells ml−1 on glass coverslips in Dulbecco’s modified Eagle’s medium, 10% FBS, GlutaMAX, 100 units/ml penicillin, and 100 μg ml−1 streptomycin. Cultures were infected with equal amount (~1 x 107 PFU) of Lenti-MCSF-Ctrl or Lenti-MCSF-PGRN at DIV 5, followed by treatment with 7PA2 conditioned medium59 (link)–60 (link) at DIV 7–9 for 48–72 h. Cells were fixed with 4% PFA and immunostained with anti-MAP2 antibody (1:300; Millipore). Surviving MAP2+ neurons were counted on a Leica DM5000B microscope. The experimenter who scored MAP2+ neurons was blinded to treatment of the cultures. For western blot analysis of cell-type specific markers, cultures were infected with equal amount of lentivirus at DIV5 and harvested at DIV7. Cells were immunoblotted with anti-GFAP (MAB3402, 1:5000, Millipore), TUJ1 (MRB-435P, 1:1000, Covance, Princeton, NJ), Iba1 (1:1000, Wako), or GAPDH (MAB374, 1:5000, Millipore).
Minami S.S., Min S.W., Krabbe G., Wang C., Zhou Y., Asgarov R., Li Y., Martens L.H., Elia L.P., Ward M.E., Mucke L., Farese RV J.r, & Gan L. (2014). Progranulin Protects against Amyloid β Deposition and Toxicity in Alzheimer’s Disease Mouse Models. Nature medicine, 20(10), 1157-1164.
+ Open protocol
+ Expand
5

Immunostaining of Mitochondrial Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
In brief, cells were fixed in 4% PFA (Sigma-Aldrich) for 15 min, followed by a 10-min permeabilization step (0.1% Triton X-100 in PBS for internal cell markers). The blocking step was performed by incubation in 2% BSA for 30 min. Cells were incubated with primary antibodies at 4°C overnight and further incubated with secondary antibodies for 1 h. For MitoTracker dye staining, cells were cultured in the normal media with 150 nM MitoTracker red (Invitrogen) for 30 min before further fixation and immunostaining. Antibodies against the following proteins were used at the indicated dilutions: CHCHD2 (HPA027407, 1:200; Sigma-Aldrich), SOX1 (4194S, 1:200; Cell Signaling Technology), NESTIN (MAB5326, 1:200; EMD Millipore), TUJ1 (MRB-435P, 1:500; Covance), mtTFA (ab119684, 1:500; Abcam), anti–mouse IgG-FITC (1:800; Sigma-Aldrich), and anti–rabbit IgG-Cy3 (1:1,000; Jackson ImmunoResearch Laboratories, Inc.). Nuclei were labeled with DAPI (Thermo Fisher Scientific). Colocalization coefficient studies were performed using ImageJ software by calculating Manders’ colocalization coefficient, which describes the amount of colocalizing pixels of GFP using pixels generated by RFP.
Zhu L., Gomez-Duran A., Saretzki G., Jin S., Tilgner K., Melguizo-Sanchis D., Anyfantis G., Al-Aama J., Vallier L., Chinnery P., Lako M, & Armstrong L. (2016). The mitochondrial protein CHCHD2 primes the differentiation potential of human induced pluripotent stem cells to neuroectodermal lineages. The Journal of Cell Biology, 215(2), 187-202.
+ Open protocol
+ Expand
6

Immunofluorescent Analysis of Neural Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cultures were fixed with 4 % paraformaldehyde for 1 h at time points when parallel cultures on MEAs exhibited high-frequency oscillations. We used the following antibodies: monoclonal Anti-Glial Fibrillary Acidic Protein (GFAP) antibody produced in mouse (G3893, Sigma-Aldrich, St. Louis, MO) and Neuronal Class III β-Tubulin Rabbit Monoclonal Antibody (MRB-435P, Covance, Princeton, NJ). Secondary antibodies were labeled with the fluorescent dyes Alexa 488 and Cy3, respectively.
Fleischer W., Theiss S., Slotta J., Holland C, & Schnitzler A. (2015). High-frequency voltage oscillations in cultured astrocytes. Physiological Reports, 3(5), e12400.
+ Open protocol
+ Expand
7

Immunofluorescence Staining of Embryoid Bodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell cultures were fixed with 4% paraformaldehyde for 20 min and permeabilized with 0.3% Triton X-100 (Sigma). Primary antibodies were incubated overnight at 4°C in phosphate buffer saline (PBS) plus 10% normal goat serum. Cells were washed three times and secondary antibodies incubated for 2 h in the same solution, washed three times and incubated with Hoechst for nuclei staining. Primary antibodies were used with the following dilutions: Oct 3/4, 1:200 (BD 611202); Sox2, 1:1000 (Millipore AB5603); GFP, 1:1000 (Invitrogen A11122); Olig2, 1:1000 (Millipore MANB50); Islet1, 1:10 (developmental studies hybridoma bank 40.3A4-5 and 40.2D6); TUJ1 (recognizing Tubulin β III), 1:1000 (Covance MRB-435P); Microtubule Associated Protein (MAP2), 1:1000 (Chemicon MAB378); Phospho Histone H3, 1:100 (Cell Signaling 9701S). Alexa Fluor secondary antibodies were used at 1:1000. EBs cultured for 5 days were harvested by gravity, washed with PBS and paraformaldehyde-fixed, cryo-protected by sucrose, tissue tek-embedded, cut in 20 μm slices and mounted in slides for immunofluorescence protocol.
Cortés D., Robledo-Arratia Y., Hernández-Martínez R., Escobedo-Ávila I., Bargas J, & Velasco I. (2016). Transgenic GDNF Positively Influences Proliferation, Differentiation, Maturation and Survival of Motor Neurons Produced from Mouse Embryonic Stem Cells. Frontiers in Cellular Neuroscience, 10, 217.
+ Open protocol
+ Expand
8

Immunostaining of Neural Progenitors and Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
All neural progenitors, including MGE cells, and further differentiated cells, including MGE cell-derived GABAergic neurons, were fixed in 4% paraformaldehyde and stained with the following primary antibodies: nestin (MAB5326, EMD Millipore), Pax6 (Developmental Studies Hybridoma Bank), FoxG1 (sc-48788, Santa Cruz Biotechnology), MAP2 (MAB3418, AB5622, EMD Millipore), Tuj1 (MAB1637, EMD Millipore; MRB-435P, Covance), apoE (178479, Calbiochem), PHF1 (gift from Peter Davies), AT8 (MN1020, Thermo Fisher Scientific), AT180 (MN1040, Thermo Fisher Scientific), Tau5 (577801, EMD Millipore), total-tau (T6402, Sigma), NKX2.1 (sc-13040, Santa Cruz Biotechnology), GABA (A2052, Sigma), and cleaved Caspase-3 (D3E9, Cell Signaling Technology). The secondary antibodies were IgG-conjugated Alexa Fluor 488 or Alexa Fluor 594 (Life Technologies). Nuclei were stained with DAPI. Images were taken with a Leica epifluorescence microscope, a Keyence BZ-9000E fluorescence microscope, or a Leica confocal imaging system.
Wang C., Najm R., Xu Q., Jeong D.E., Walker D., Balestra M.E., Yoon S.Y., Yuan H., Li G., Miller Z.A., Miller B.L., Malloy M.J, & Huang Y. (2018). Gain of toxic Apolipoprotein E4 effects in Human iPSC-Derived Neurons Is Ameliorated by a Small-Molecule Structure Corrector. Nature medicine, 24(5), 647-657.
+ Open protocol
+ Expand
9

Immunocytochemical Staining and Quantification of Radial Glia in Hippocampus

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunocytochemicalstaining was performed as previously described (Sessa et al., 2019 (link)).
The primary antibodies utilized were the following: anti-KI67 (Immunological Science, MAB90948), anti-GFAP (Merck, AB5804), anti-TUJ1 (Covance, MRB-435P), anti-phospho histone 3 (PH3) (rabbit, 1:400, Sigma-Aldrich, H0412), anti-S100b (Dako, GA504). Anti-O4 primary antibody was produced from a hybridoma clone, using the culture media of hybridoma cells directly on living cells to perform the staining for O4 epitope. Secondary antibody used: 488-mouse (donkey, 1:2,000, Molecular Probes, A21202), 488-rabbit (donkey, 1:2,000, Molecular Probes, A21026), 594-mouse (donkey, 1:2,000, Molecular Probes, A10036), 594-rabbit (donkey, 1:2,000, Molecular Probes, A21207). DAPI (4′,6′-diamidino-2-phenylindole) was used as nuclear counterstaining.
The quantification of Radial Glia cells in the hippocampus has been performed using GFAP antibody, counting only the GFAP+ cells present in the subgranular zone of the dentate gyrus.
Mastrototaro G., Zaghi M., Massimino L., Moneta M., Mohammadi N., Banfi F., Bellini E., Indrigo M., Fagnocchi G., Bagliani A., Taverna S., Rohm M., Herzig S, & Sessa A. (2021). TBL1XR1 Ensures Balanced Neural Development Through NCOR Complex-Mediated Regulation of the MAPK Pathway. Frontiers in Cell and Developmental Biology, 9, 641410.
+ Open protocol
+ Expand
10

Immunostaining of Neural Progenitors and Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
All neural progenitors, including MGE cells, and further differentiated cells, including MGE cell-derived GABAergic neurons, were fixed in 4% paraformaldehyde and stained with the following primary antibodies: nestin (MAB5326, EMD Millipore), Pax6 (Developmental Studies Hybridoma Bank), FoxG1 (sc-48788, Santa Cruz Biotechnology), MAP2 (MAB3418, AB5622, EMD Millipore), Tuj1 (MAB1637, EMD Millipore; MRB-435P, Covance), apoE (178479, Calbiochem), PHF1 (gift from Peter Davies), AT8 (MN1020, Thermo Fisher Scientific), AT180 (MN1040, Thermo Fisher Scientific), Tau5 (577801, EMD Millipore), total-tau (T6402, Sigma), NKX2.1 (sc-13040, Santa Cruz Biotechnology), GABA (A2052, Sigma), and cleaved Caspase-3 (D3E9, Cell Signaling Technology). The secondary antibodies were IgG-conjugated Alexa Fluor 488 or Alexa Fluor 594 (Life Technologies). Nuclei were stained with DAPI. Images were taken with a Leica epifluorescence microscope, a Keyence BZ-9000E fluorescence microscope, or a Leica confocal imaging system.
Wang C., Najm R., Xu Q., Jeong D.E., Walker D., Balestra M.E., Yoon S.Y., Yuan H., Li G., Miller Z.A., Miller B.L., Malloy M.J, & Huang Y. (2018). Gain of toxic Apolipoprotein E4 effects in Human iPSC-Derived Neurons Is Ameliorated by a Small-Molecule Structure Corrector. Nature medicine, 24(5), 647-657.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!