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Nb100 60554

Manufactured by Novus Biologicals
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Sourced in United Kingdom

The NB100-60554 is a laboratory equipment product from Novus Biologicals. It is designed for general laboratory use. The core function of this product is to provide a reliable and consistent platform for various laboratory procedures and experiments.

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Lab products found in correlation

Axioobserver epifluorescence microscope, by Zeiss (5 mentions) Ab10983, by Abcam (4 mentions) Alexa fluor 488 goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions) Fsx100 microscope, by Olympus (2 mentions) Alexa fluor 488 donkey anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 donkey anti goat igg h l, by Thermo Fisher Scientific (1 mentions) Sc 59171, by Santa Cruz Biotechnology (1 mentions) Axio observer 7, by Zeiss (1 mentions) Dapi containing mounting media, by Thermo Fisher Scientific (1 mentions) Ab124432, by Abcam (1 mentions)

7 protocols using nb100 60554

1

Immunostaining of Adipose Tissue Markers

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Formalin-fixed, paraffin-embedded sections from adipose tissue (n = 2–3 male mice) were blocked in PBST with 5% BSA. Primary antibody used was perilipin (1:500 dilution) (a kind gift of Dr. Andy Greenberg, Tufts University or NB100-60554, NOVUS) and UCP1 (1:250 dilution) (ab10983, Abcam, Cambridge, England); secondary antibodies (1:200 dilution) used were Alexa Fluor 594 Goat anti-Rabbit IgG (H+L), Alexa Fluor 594 Donkey anti-Goat IgG (H+L) and Alexa Fluor 488 Goat anti-Rabbit IgG (H+L) (Invitrogen, Carlsbad, CA). Slides were counterstained with DAPI. Images were acquired using AxioObserver Epifluorescence Microscope (Zeiss, Jena, Germany).
Wang Q.A., Tao C., Jiang L., Shao M., Ye R., Zhu Y., Gordillo R., Ali A., Lian Y., Holland W.L., Gupta R.K, & Scherer P.E. (2015). Distinct Regulatory Mechanisms Govern Embryonic versus Adult Adipocyte Maturation. Nature cell biology, 17(9), 1099-1111.
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2

Imaging Adipose Tissue Thermogenesis

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Formalin‐fixed, paraffin‐embedded sections from adipose tissue (n = 2–3 male mice) were blocked in PBST with 5% BSA (Wang et al, 2015). Primary antibody used was perilipin (1:500 dilution) (NB100‐60554, NOVUS) and UCP1 (1:250 dilution) (ab10983, Abcam, Cambridge, England); secondary antibodies (1:200 dilution) used were Alexa Fluor 594 Donkey anti‐Goat IgG (H+L) and Alexa Fluor 488 Donkey anti‐Rabbit IgG (H+L) (Invitrogen, Carlsbad, CA, USA). Slides were counterstained with DAPI. Images were acquired using AxioObserver Epifluorescence Microscope (Zeiss, Jena, Germany).
Wang Z., Ning T., Song A., Rutter J., Wang Q.A, & Jiang L. (2020). Chronic cold exposure enhances glucose oxidation in brown adipose tissue. EMBO Reports, 21(11), e50085.
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3

Immunostaining of Adipose Tissue Markers

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Formalin-fixed, paraffin-embedded sections from adipose tissue (n = 2–3 male mice) were blocked in PBST with 5% BSA. Primary antibody used was perilipin (1:500 dilution) (a kind gift of Dr. Andy Greenberg, Tufts University or NB100-60554, NOVUS) and UCP1 (1:250 dilution) (ab10983, Abcam, Cambridge, England); secondary antibodies (1:200 dilution) used were Alexa Fluor 594 Goat anti-Rabbit IgG (H+L), Alexa Fluor 594 Donkey anti-Goat IgG (H+L) and Alexa Fluor 488 Goat anti-Rabbit IgG (H+L) (Invitrogen, Carlsbad, CA). Slides were counterstained with DAPI. Images were acquired using AxioObserver Epifluorescence Microscope (Zeiss, Jena, Germany).
Wang Q.A., Tao C., Jiang L., Shao M., Ye R., Zhu Y., Gordillo R., Ali A., Lian Y., Holland W.L., Gupta R.K, & Scherer P.E. (2015). Distinct Regulatory Mechanisms Govern Embryonic versus Adult Adipocyte Maturation. Nature cell biology, 17(9), 1099-1111.
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4

Quantifying Adipose Tissue Fibrosis and CLSs

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Adipose fat tissues and liver were fixed in 4% paraformaldehyde and embedded in paraffin. Areas of fibrosis were measured using image analysis software (Axio Observer 7, ZEISS). CLSs in Epi-fat were detected immunohistochemically using an anti-F4/80 antibody (sc-59171, Santa Cruz Biotechnology) and an anti-perilipin antibody (NB100-60554, Novus Biologicals). The CLS density was obtained from the number of CLSs per 1,000 adipocytes in each section45 (link). They were counted in more than 5 mm2 areas of each section per mouse, and the number of CLSs was expressed as the mean number per mm2.
Nakazeki F., Nishiga M., Horie T., Nishi H., Nakashima Y., Baba O., Kuwabara Y., Nishino T., Nakao T., Ide Y., Koyama S., Kimura M., Tsuji S., Sowa N., Yoshida S., Conway S.J., Yanagita M., Kimura T, & Ono K. (2018). Loss of periostin ameliorates adipose tissue inflammation and fibrosis in vivo. Scientific Reports, 8, 8553.
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5

Immunofluorescence Analysis of Adipose Tissue

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Immunofluorescence was done as previously described [29 (link)]. In brief, formalin-fixed, paraffin-embedded sections from white adipose and brown adipose tissues were blocked in PBST with 5% BSA. Primary antibodies used were against perilipin (1:500 dilution NB100-60554, Novus), UCP1 (1:250 dilution, ab10983, Abcam), or Mac2 (1:500 dilution, Cat#: 125,401, BioLegend). Secondary antibodies (1:250 dilution) used were Alexa Fluor 488 or 594 donkey anti-rabbit IgG (HCL) or Alexa Fluor 488 or 594 donkey anti-goat IgG (HCL) (Invitrogen). Slides were counterstained with DAPI. Fluorescent images were acquired using an AxioObserver Epifluorescence Microscope (Zeiss) or an FSX100 microscope (Olympus).
Zhao S., Li N., Zhu Y., Straub L., Zhang Z., Wang M.Y., Zhu Q., Kusminski C.M., Elmquist J.K, & Scherer P.E. (2020). Partial leptin deficiency confers resistance to diet-induced obesity in mice. Molecular Metabolism, 37, 100995.
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6

FISH-based Satellite Heterochromatin Imaging

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SADS assay, which is a fluorescence in situ hybridization (FISH) for detection of peri-centromeric satellite heterochromatin DNA, was performed following the protocol kindly provided by Drs. Sundeep Khosla and Joshua Farr76 (link). Briefly, paraffin-embedded femoral bone tissue sections were cross-linked with 4% paraformaldehyde and dehydrated in ethanol. The bone tissue sections were then denatured and hybridized in the buffer containing 1.0 μg/mL Cy3-labeled, CENPB-specific (ATTCGTTGGAAACGGGA) peptide nucleic acid FISH probe (Panagene Inc, Korea) for 2 hours at room temperature in the dark. After washing, immunofluorescence staining of the tissue sections was performed using primary antibody against perilipin (Novus Biologicals, NB100-60554, 1:500). Finally, the slides were mounted with DAPI-containing mounting media (Life Technologies). Images were acquired with a Zeiss LSM780 confocal microscope, and the intensity profiles were acquired with Image J software (NIH).
Lemaire V., Koehl M., Le Moal M, & Abrous D.N. (2000). Prenatal stress produces learning deficits associated with an inhibition of neurogenesis in the hippocampus. Proceedings of the National Academy of Sciences of the United States of America, 97(20), 11032-11037.
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7

Adipose and Liver Immunofluorescence

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Immunofluorescence was performed as previously described (45) . Briefly, formalin-fixed, paraffin-embedded sections from adipose tissues or liver were blocked in 1× phosphate-buffered saline with 0.1% Tween 20 (PBST) containing 5% bovine serum albumin. Primary antibodies used were perilipin (1:500 dilution; #NB100-60554, Novus; RRID: AB_922242), CD31 (1:250 dilution; #ab124432, Abcam; RRID: N/A), and Mac2 (1:500 dilution; #125401, BioLegend; RRID: AB_1134237). Secondary antibodies (1:250 dilution) used were Alexa Fluor 488 or Alexa Fluor 594 donkey anti-rabbit IgG both heavy and light (H+L) chains or Alexa Fluor 488 or Alexa Fluor 594 donkey anti-goat IgG (H+L) (Thermo Fisher Scientific). Slides were counterstained with 4 0 ,6-diamidino-2-phenylindole. Fluorescent images were acquired using an AxioObserver Epifluorescence Microscope (Zeiss) or FSX100 microscope (Olympus).
Zhao S., Lin Q., Xiong W., Li L., Straub L., Zhang D., Zapata R., Zhu Q., Sun X.N., Zhang Z., Funcke J.B., Li C., Chen S., Zhu Y., Jiang N., Li G., Xu Z., Wyler S.C., Wang M.Y., Bai J., Han X., Kusminski C.M., Zhang N., An Z., Elmquist J.K., Osborn O., Liu C, & Scherer P.E. (2023). Hyperleptinemia contributes to antipsychotic drug-associated obesity and metabolic disorders. Science translational medicine, 15(723).
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