The following antibodies K19 (A53-B/A2), GSK3β (E-11), pGSK3β (F-2), PARP-1 (F-2), β-actin (C4), and GAPDH (0411) were from Santa Cruz Biotechnology (Santa Cruz, CA); cyclin D3 (DCS22) and GSK3β (D5C5Z) were from Cell Signaling Technology (Danvers, MA); GFP (3H9) and RFP (5F8) were from Chromotek (Islandia, NY); cyclin D3 (26755-1-AP) was from Proteintech (Rosemont, IL); and anti-tubulin (12G1), anti-GFP (1D2), and c-Myc (9E10) were from Developmental Studies Hybridoma Bank (Iowa City, IA). Forskolin was from ApexBio (#B1421) (Houston, TX), LiCl (# EM-LX0331) and DMSO were from VWR Life Sciences (Carlsbad, CA), LY294002 was from AmBeed (#A133122) (Arlington Hts, IL), SB 203580 (#S-3400) and SB 202190 (S-1700) were from LC Laboratories (Woburn, MA), okadaic acid (459616) was from EMD Millipore (Billerica, MA), cycloheximide (#66-81-9) was from Sigma-Aldrich (St Louis, MO), and MG132 (S2619) was from Selleck Chemicals.
Cyclin d3 dcs22
Manufactured by Cell Signaling Technology
Cyclin D3 (DCS22) is a primary antibody used in immunodetection applications. Cyclins are a family of proteins that play a crucial role in regulating the cell cycle. Cyclin D3 is specifically involved in the G1/S transition of the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Jak3 d7b12, by Cell Signaling Technology (2 mentions)
Hrp linked goat anti rabbitsecondary, by Southern Biotech (2 mentions)
Odyssey block, by LI COR (2 mentions)
Odyssey scanner, by LI COR (2 mentions)
Chemidoc mp, by Bio-Rad (2 mentions)
Mg132 s2619, by Selleck Chemicals (1 mentions)
Gapdh 0411, by Santa Cruz Biotechnology (1 mentions)
Rfp 5f8, by Proteintech (1 mentions)
C myc 9e10, by Developmental Studies Hybridoma Bank (1 mentions)
Forskolin, by Apexbio (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Sb203580, by LC Laboratories (1 mentions)
Hematoxylin, by Thermo Fisher Scientific (1 mentions)
Ki 67 antibody ab15580, by Abcam (1 mentions)
Dab reaction, by Vector Laboratories (1 mentions)
Abc substrate kit, by Vector Laboratories (1 mentions)
Prb s780, by Cell Signaling Technology (1 mentions)
4 protocols using cyclin d3 dcs22
1
Comprehensive Antibody and Reagent Source
2
Western Blotting of Cyclin D3 and JAK3
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5–10 × 106 cells were lysed in 50 μL
RIPA buffer with 500 mM NaCl, clarified, and protein normalized by bicinchoninic
acid assay (Pierce). Proteins (20–30 μg) were resolved by
7.5% or 10% SDS-PAGE, transferred to a polyvinylidene fluoride
membrane and blocked with Odyssey Block (LI-COR Biosciences). Blots were probed
for Cyclin D3 (DCS22, 1:1000) or JAK3 (D7B12, 1:1000) (Cell Signaling
Technologies), normalized to tubulin (DM1A, 1:5000, Sigma-Aldrich) and detected
with fluorescent secondary antibodies on an Odyssey Scanner (LI-COR
Biosciences). Alternatively, blots were probed with HRP-linked goat-anti-rabbit
secondary (Southern Biotech, Birmingham AL), developed with Western Lightening
Plus ECL (Pierce) and detected on a ChemiDocMP (Biorad).
RIPA buffer with 500 mM NaCl, clarified, and protein normalized by bicinchoninic
acid assay (Pierce). Proteins (20–30 μg) were resolved by
7.5% or 10% SDS-PAGE, transferred to a polyvinylidene fluoride
membrane and blocked with Odyssey Block (LI-COR Biosciences). Blots were probed
for Cyclin D3 (DCS22, 1:1000) or JAK3 (D7B12, 1:1000) (Cell Signaling
Technologies), normalized to tubulin (DM1A, 1:5000, Sigma-Aldrich) and detected
with fluorescent secondary antibodies on an Odyssey Scanner (LI-COR
Biosciences). Alternatively, blots were probed with HRP-linked goat-anti-rabbit
secondary (Southern Biotech, Birmingham AL), developed with Western Lightening
Plus ECL (Pierce) and detected on a ChemiDocMP (Biorad).
3
Immunohistochemical Analysis of Lymphoma Xenografts
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Lymphoma tissues from xenograft were fixed with 4%PFA, dehydrated, and embedded in paraffin followed by sectioned with a microtome. H&E sections and tissue microarray slides (US Biomax, Inc.) were blocked with 10% goat serum, incubated with the primary antibodies and subsequently incubated with biotinylated antibodies. Signal was developed with ABC substrate kit (Vector) followed by DAB reaction (Vector), and counterstained with Hematoxylin (Thermo Scientific). The Ki-67 antibody (ab15580) was purchased from Abcam. Fbxl8 antibody (NBP2-34012) was obtained from Novus Biologicals. Cyclin D3 (DCS22) and pRb (S780) were purchased from Cell Signaling Technology and Santa Cruz, respectively. The IHC score in each core was defined by following formula Intensity = (staining positive population (1 to 3) × staining intensity (1 to3)). The IHC scores 1–2, 3–5, and 6–9 were defined as expression of low, medium, and high, respectively.
4
Western Blotting of Cyclin D3 and JAK3
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