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Nb600 308

Manufactured by Merck Group
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The NB600-308 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use, providing a core function of sample handling and processing. The description of the product's intended use or specific applications is not available.

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Lab products found in correlation

A11004, by Thermo Fisher Scientific (2 mentions) Fluorogold, by Southern Biotech (2 mentions) Ab144p, by Merck Group (2 mentions) Ultra emccd camera, by Oxford Instruments (1 mentions) L 15 medium, by Thermo Fisher Scientific (1 mentions) Accuri c6, by BD (1 mentions) X tremegene 9, by Roche (1 mentions)

4 protocols using nb600 308

1

Immunohistochemical Labeling Protocol for Brain Tissue

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Mice were first anesthetized and then transcardially perfused with cold 4% paraformaldehyde (PFA) in PBS (pH 7.4). Brains were fixed in 4% PFA. 50 μm coronal slices were sectioned on a vibratome, and then were subsequently stored in cryoprotectant at 4°C. For immunohistochemistry, individual sections were washed in PBS and then incubated for 30 min in 0.3% Triton-X and 3% normal donkey serum (NDS).
For ChAT labeling (Figure S6), ChAT primary antibody (1:200; Millipore, Product# AB144P [RRID: AB_2079751]) incubations were performed overnight at 4°C in 3% NDS/PBS. Sections were washed and left to incubate in secondary antibodies conjugated to AlexaFluor586 for 3 h at room temperature (1:1000; Invitrogen, Product#A-11004 [RRID: AB_10564097]).
For enhancement of GFP in GACh3.0 tissue labeling (Figure 2B), GFP primary antibody (1:500; Millipore, product # NB600-308 [RRID: AB_10003058]) incubations were performed overnight at 4°C in 3% NDS/PBS. Sections were washed and left to incubate in secondary antibodies conjugated to AlexaFluor586 for 3 h at room temperature (1:1000; Invitrogen, Product#A-11004 [RRID: AB_10564097]).
Following a 20 min incubation with DAPI (1:50,000) sections were washed and mounted on microscope slides with Fluorogold (Southern Biotechnology, product # 0100-01).
Fleming W., Lee J., Briones B.A., Bolkan S.S, & Witten I.B. (2022). Cholinergic interneurons mediate cocaine extinction in male mice through plasticity across medium spiny neuron subtypes. Cell reports, 39(9), 110874.
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2

Immunohistochemical Labeling Protocol for Brain Tissue

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Mice were first anesthetized and then transcardially perfused with cold 4% paraformaldehyde (PFA) in PBS (pH 7.4). Brains were fixed in 4% PFA. 50 μm coronal slices were sectioned on a vibratome, and then were subsequently stored in cryoprotectant at 4°C. For immunohistochemistry, individual sections were washed in PBS and then incubated for 30 min in 0.3% Triton-X and 3% normal donkey serum (NDS).
For ChAT labeling (Figure S6), ChAT primary antibody (1:200; Millipore, Product# AB144P [RRID: AB_2079751]) incubations were performed overnight at 4°C in 3% NDS/PBS. Sections were washed and left to incubate in secondary antibodies conjugated to AlexaFluor586 for 3 h at room temperature (1:1000; Invitrogen, Product#A-11004 [RRID: AB_10564097]).
For enhancement of GFP in GACh3.0 tissue labeling (Figure 2B), GFP primary antibody (1:500; Millipore, product # NB600-308 [RRID: AB_10003058]) incubations were performed overnight at 4°C in 3% NDS/PBS. Sections were washed and left to incubate in secondary antibodies conjugated to AlexaFluor586 for 3 h at room temperature (1:1000; Invitrogen, Product#A-11004 [RRID: AB_10564097]).
Following a 20 min incubation with DAPI (1:50,000) sections were washed and mounted on microscope slides with Fluorogold (Southern Biotechnology, product # 0100-01).
Fleming W., Lee J., Briones B.A., Bolkan S.S, & Witten I.B. (2022). Cholinergic interneurons mediate cocaine extinction in male mice through plasticity across medium spiny neuron subtypes. Cell reports, 39(9), 110874.
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3

Antibodies Used in Cellular Assays

Cited 2 times
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The following antibodies were used in this study: anti-MISP (rb)2 (link) anti-MISP (ms, kind gift from Alwin Krämer)3 (link), anti-IQGAP1 (rb, Abcam 86064), anti-IQGAP1 (ms, Santa Cruz sc-376021), anti-Cdc42 (ms, Cytoskeleton, ACD03), anti-pericentrin (rb, Abcam ab4448), anti-β-actin (ms, Calbiochem JLA20), anti-α-tubulin (ms, Sigma B-5-1-2, T5168), anti-GFP (rb, Novus Biologicals NB600-308), anti-FLAG M2 (ms, Sigma F1804), anti-GST Z5 (rb, Santa Cruz sc-459), anti-EB1 (ms, BD Biosciences 610535), anti-pAkt (S473, ms, CST #587F11), anti-Akt (rb, CST #9272), anti-p150glued (ms, BD Biosciences 612709). Secondary antibodies: Alexa 488/568/647-coupled anti-rabbit/mouse IgG (Molecular probes).
Vodicska B., Cerikan B., Schiebel E, & Hoffmann I. (2018). MISP regulates the IQGAP1/Cdc42 complex to collectively orchestrate spindle orientation and mitotic progression. Scientific Reports, 8, 6330.
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4

Quantifying TDP-43 Expression and Aggregation

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Mammalian TDP‐43 expression constructs (500 ng) were transfected into HEK‐293T cells grown on tissue culture‐treated μ‐slides (ibidi) with X‐tremeGENE 9 (Roche). To compare expression levels, cells were harvested 24 h after transfection and analyzed by (i) flow cytometry with a BD Accuri C6 instrument (excitation laser: 488 nm, emission filter: 586/40 nm) or (ii) immunoblotting using anti‐GFP (Novus NB600‐308) and anti‐alpha‐tubulin (Sigma T6199) primary antibodies. Blots were developed with a LiCor Odyssey imager using appropriate dye‐conjugated secondary antibodies. To follow TDP‐43 reporter phase formation over time, cells were imaged in L15 medium (Gibco) + 10% FBS at 37°C every hour for a total of 24 h after transfection with a Leica DMI‐6000B microscope equipped with an Andor Yokogawa spinning disk confocal unit. A 488‐nm laser was used for GFP excitation and an Andor iXon Ultra EMCCD camera for signal detection.
Wang A., Conicella A.E., Schmidt H.B., Martin E.W., Rhoads S.N., Reeb A.N., Nourse A., Ramirez Montero D., Ryan V.H., Rohatgi R., Shewmaker F., Naik M.T., Mittag T., Ayala Y.M, & Fawzi N.L. (2018). A single N‐terminal phosphomimic disrupts TDP‐43 polymerization, phase separation, and RNA splicing. The EMBO Journal, 37(5), e97452.
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