Anti IgG Fc  

ERK is required for fibrous tissue marker regulation but not required to inhibit chondrogenesis. (A) Sclerotome was treated with a MEK inhibitor PD184352, PD, to inhibit ERK activity, for 24？h and then cells were treated with TGFβ1 for 8？h. Immunoblot was used to determine relative levels of pERK1/2, ERK1/2, and α-tubulin. Primary antibodies: pERK1/2 , ERK1/2, and Alpha tubulin [p/n 200-301-880] with secondary antibodies: anti-Rabbit-HRP and anti-mouse DyLight？649 [p/n 610-143-003]. Figure 5. PMID: 33288795.

TGF-β signaling regulates noncanonical pathways in the sclerotome. Sclerotome was treated with vehicle control or TGFβ1 for 2, 8 or 24？h. Immunoblot was used to determine activity of ERK, p38 and AKT. α tubulin was used as a general loading control. Immunoblots were cropped for clarity. Primary antibodies: pERK1/2 , ERK1/2, pp38, p38, pAKT, AKT, and Alpha tubulin [p/n 200-301-880] with secondary antibodies: anti-Rabbit-HRP and anti-mouse DyLight？649 [p/n 610-143-003]. Figure 3. PMID: 33288795.

Knock-down of？Sox9？attenuates TGF-β-mediated regulation of？Papss2 ATDC5 cells were transfected with？Sox9？siRNA,？Smad2/3？siRNA, or control non-specific (NS) siRNA.？Western blots showed Smad2/3 protein levels were reduced in the presence of？Smad2/3？siRNA, α-Tubulin was used as a loading control (n = 6) (D).？Primary antibodies: anti-SMAD2/3 antibody (1:2000) or anti-α-Tubulin antibody [1:2500, p/n 200-301-880], with secondary antibodies anti-mouse [1:2500, p/n 610-143-003]. Figure 4. PMID: 27746378.

SMAD3 and SOX9 adenoviral vectors infect cells, induce expression of FLAG-tagged proteins, and up-regulate SMAD3 and SOX9 function respectively Bovine chondrocytes that were infected with either Ad-eGFP, Ad-SMAD3, or Ad-SOX9. Western blots of extracts from bovine chondrocytes showed expression of FLAG-tagged proteins at approximately 52 kDa and 75 kDa, corresponding to SMAD3 and SOX9 molecular weights respectively (n = 4) (B). Primary antibodies: anti-GAPDH antibody (1:1000) or anti-FLAG antibody (1:500), with secondary antibodies: HRP-conjugated anti-rabbit (1:2000) and anti-mouse [1:2500, p/n 610-143-003] Figure 2. PMID: 27746378.

DyLight？ dyes can be used for two-color Western Blot detection with low background and high signal.？ Anti-tubulin was detected using a DyLight？ 549 conjugate.？ Anti-TNFa was detected using a DyLight？ 649 conjugate. The image was captured using the Typhoon？ 9410 Imaging System.

Properties of DyLight？ Conjugates.