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D2944

Manufactured by Merck Group
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D2944 is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It is a versatile instrument capable of performing a wide range of chromatographic separations, including reversed-phase, normal-phase, and ion-exchange chromatography. The core function of the D2944 is to facilitate the separation, identification, and quantification of complex mixtures of chemical compounds.

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Lab products found in correlation

Ab5084p, by Merck Group (2 mentions) Donkey anti rat alexa 488, by Thermo Fisher Scientific (2 mentions) Donkey anti mouse alexa 488, by Thermo Fisher Scientific (2 mentions) Rat anti d1, by Merck Group (2 mentions) Mouse anti d2, by Santa Cruz Biotechnology (2 mentions) Supersignal west pico plus, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Ibright fl1000, by Thermo Fisher Scientific (1 mentions) Am4302, by Thermo Fisher Scientific (1 mentions) Modified bradford assay, by Bio-Rad (1 mentions) Gapdh, by Merck Group (1 mentions) Mab318, by Merck Group (1 mentions) Sc 48, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Sigma-D2944 (2 citations)

7 protocols using d2944

1

Protein Isolation and Western Blot Analysis

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Total protein was isolated and quantified using a modified Bradford assay (Bio‐Rad). Western blot analyses were performed using the following primary antibodies: mouse anti‐COASY (Santa Cruz sc‐393812, 1/1,000), rat anti‐D1 dopamine receptor (Sigma D2944, 1/1,000), mouse anti‐TFR1 (ThermoFisher 13‐6800, 1/3,000), rabbit anti‐pyruvate dehydrogenase (Cell Signaling 3205, 1/3,000), rabbit anti‐VDAC (Cell Signaling 4661, 1/5,000), mouse anti‐MSH6 (BD 610918, 1/10,000), and mouse anti‐β actin (ThermoFisher AM4302, 1/10,000). Most of the primary antibody binding was visualized with the traditional method using a HRP‐conjugated secondary antibody (Jackson ImmunoResearch, 1/10,000) and ECL substrate (SuperSignal™ West Pico PLUS, ThermoFisher). However, anti‐DRD1 and its loading control (anti‐MSH6) were visualized using a biotinylated secondary antibody and Alexa Fluor™ 488‐conjugated streptavidin (ThermoFisher, 1/4,000). The fluorescent Western blot signal was captured using iBright™ FL1000 (ThermoFisher).
Jeong S.Y., Hogarth P., Placzek A., Gregory A.M., Fox R., Zhen D., Hamada J., van der Zwaag M., Lambrechts R., Jin H., Nilsen A., Cobb J., Pham T., Gray N., Ralle M., Duffy M., Schwanemann L., Rai P., Freed A., Wakeman K., Woltjer R.L., Sibon O.C, & Hayflick S.J. (2019). 4′‐Phosphopantetheine corrects CoA, iron, and dopamine metabolic defects in mammalian models of PKAN. EMBO Molecular Medicine, 11(12), e10489.
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2

Dopamine Receptor Protein Quantification

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The primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit).
Hasbi A., Perreault M.L., Shen M.Y., Fan T., Nguyen T., Alijaniaram M., Banasikowski T.J., Grace A.A., O'Dowd B.F., Fletcher P.J, & George S.R. (2018). Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB. Frontiers in Pharmacology, 8, 924.
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3

Immunohistochemical Analysis of Dopaminergic Markers

Cited 2 times
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The immunohistochemistry was performed in the adult behaviorally naïve mice using DAB immunostaining. The following primary antibodies were used: rabbit anti-tyrosine hydroxylase (TH) (Santa Cruz Biotechnology sc14007; diluted 1:500), rat anti dopamine transporter (DAT) (Chemicon MAB369; diluted 1:1000), rat anti-dopamine D1 receptor (D1R) (Sigma Aldrich D2944; diluted 1:1000) and rabbit anti-dopamine D2 receptor (D2R) (Chemicon AB5084P; diluted 1:500). The specificity of immunoreactions of the primary antibodies were assessed and validated in several recent publications14 (link),27 (link),28 (link). Microscopic analysis and quantification were conducted using densitometry and steriological methods. The full description of the tissue processing for the immunohistochemistry and the image quantification methods are provided in the supplementary file.
Sarker G., Berrens R., von Arx J., Pelczar P., Reik W., Wolfrum C, & Peleg-Raibstein D. (2018). Transgenerational transmission of hedonic behaviors and metabolic phenotypes induced by maternal overnutrition. Translational Psychiatry, 8, 195.
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4

Immunohistochemical Detection of Dopamine Receptors

Cited 1 time
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A second and third set of tissues was assayed for D1 and D2. For D1, tissues were incubated in primary antibody rat anti-D1 (1:1,000, Sigma Aldrich D2944) for 48 hours at 4°C. Specificity of D1 antibody was validated by omission of primary antibody and by a previous prebsorption and Western blot study (24 (link)). Tissues were incubated for 1 hour in donkey anti-rat/Alexa 488 (1:1,000, Life Technologies A21208) and counterstained with DAPI. For D2, tissues underwent antigen retrieval for 30 minutes in 10 mM sodium citrate buffer (pH 8.5) to unmask immunosignal and were incubated in primary antibody mouse anti-D2 (1:200, Santa Cruz sc-5303) for 48 hours at 4°C. Specificity of D2 antibody was validated by omission of primary antibody and by previous preabsorption and Western blot studies (25 (link), 26 (link)). Tissues were incubated for 1 hour in donkey anti-mouse/Alexa 488 (1:1,000, Life Technologies A21202).
Rivera H.M., Kievit P., Kirigiti M.A., Bauman L.A., Baquero K., Blundell P., Dean T.A., Valleau J.C., Takahashi D.L., Frazee T., Douville L., Majer J., Smith M.S., Grove K.L, & Sullivan E.L. (2015). Maternal High-Fat Diet and Obesity Impact Palatable Food Intake and Dopamine Signaling in Nonhuman Primate Offspring. Obesity (Silver Spring, Md.), 23(11), 2157-2164.
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5

Immunohistochemical Detection of Dopamine Receptors

Cited 1 time
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A second and third set of tissues was assayed for D1 and D2. For D1, tissues were incubated in primary antibody rat anti-D1 (1:1,000, Sigma Aldrich D2944) for 48 hours at 4°C. Specificity of D1 antibody was validated by omission of primary antibody and by a previous prebsorption and Western blot study (24 (link)). Tissues were incubated for 1 hour in donkey anti-rat/Alexa 488 (1:1,000, Life Technologies A21208) and counterstained with DAPI. For D2, tissues underwent antigen retrieval for 30 minutes in 10 mM sodium citrate buffer (pH 8.5) to unmask immunosignal and were incubated in primary antibody mouse anti-D2 (1:200, Santa Cruz sc-5303) for 48 hours at 4°C. Specificity of D2 antibody was validated by omission of primary antibody and by previous preabsorption and Western blot studies (25 (link), 26 (link)). Tissues were incubated for 1 hour in donkey anti-mouse/Alexa 488 (1:1,000, Life Technologies A21202).
Rivera H.M., Kievit P., Kirigiti M.A., Bauman L.A., Baquero K., Blundell P., Dean T.A., Valleau J.C., Takahashi D.L., Frazee T., Douville L., Majer J., Smith M.S., Grove K.L, & Sullivan E.L. (2015). Maternal High-Fat Diet and Obesity Impact Palatable Food Intake and Dopamine Signaling in Nonhuman Primate Offspring. Obesity (Silver Spring, Md.), 23(11), 2157-2164.
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6

Immunohistochemical Mapping of Dopaminergic Pathways

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Coronal rat brain sections (50 µm) were collected and processed for tyrosine hydroxylase (MAB318, Milipore), ∆FosB (sc-48, Santa-Cruz) and D1R (D2944, Sigma) as previously described5 (link), 8 (link), 9 (link), 40 .
Bastide M.F., Glangetas C., Doudnikoff E., Li Q., Bourdenx M., Fernagut P.O., Dumont É.C., Georges F, & Bézard E. (2017). Involvement of the bed nucleus of the stria terminalis in L-Dopa induced dyskinesia. Scientific Reports, 7, 2348.
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7

Direct Fluorescent Labeling of D1R and D2R

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For experiments of D1R and D2R colocalization, we eliminated any use of secondary antibodies by directly conjugating the anti-D1R antibody (Sigma, D2944) to Alexa Fluor-488, and the anti-D2R antibody (Millipore, AB5084P) to Alexa Fluor-568, accordingly to the manufacturer's instructions (Invitrogen). Coronal sections (16 μm, Bregma:1.6 ± 0.6) from perfused rat brain were blocked (1% BSA, 0.1% Triton-X in 0.05M PBS) and then incubated for 96 h at 4°C with anti-D1-488 (1:200) and anti-D2-568 (1:200). After three washes, the last one containing DAPI, the slides were mounted (Dako), and the images were acquired in sequential mode in the same conditions described above.
Hasbi A., Perreault M.L., Shen M.Y., Fan T., Nguyen T., Alijaniaram M., Banasikowski T.J., Grace A.A., O'Dowd B.F., Fletcher P.J, & George S.R. (2018). Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB. Frontiers in Pharmacology, 8, 924.
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