To induce neurogenic differentiation, fifth-passage BM-MSCsPRL−1 were seeded at a density of 5 × 103 cells/cm2 using a multistep protocol. After 12 days, neurogenic differentiation was confirmed by immunofluorescence analysis of glial fibrillary acidic protein (GFAP) and nestin (NES). Cells fixed using 4% PFA were stained with polyclonal antibodies (1:200 dilution; Sigma-Aldrich) at 4°C overnight. An Alexa 488-conjugated polyclonal antibody (Invitrogen) was used as the secondary antibody, and DAPI was used for counterstaining (Vector Laboratories, Burlingame, CA, USA).
Polyclonal antibodies
Manufactured by Merck Group
Sourced in United States
Polyclonal antibodies are a type of laboratory-produced antibodies that are derived from multiple B cells. They recognize multiple epitopes on an antigen and are commonly used for various research and diagnostic applications.
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Lab products found in correlation
4 protocols using polyclonal antibodies
1
Neurogenic Differentiation of BM-MSCs
2
Silencing ATG7 and BECLIN1 in Cancer Cells
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Lentiviral shRNA (ATG7 and BECLIN1) constructs were purchased from Open Biosystems (RMM4534_019584 and RMM4534_028835) (Ottawa, ON, Canada). ATG7 and BECLIN1 shRNAs were transfected into PC3, LNCaP, or MCF7 cells either alone or together with packaging plasmids following the manufacturer's protocol (Invitrogen ViraPower Lentiviral Expression Systems kit, Carlsbad, CA). Cells were passaged and plated in a six-well plate and allowed to adhere for 24 hours before infection. The silencing efficacy of the various shRNAs was assessed by Western blot analysis of ATG7 and BECLIN1 proteins using polyclonal antibodies (Sigma-Aldrich, Oakville, ON, Canada).
3
Expression and Purification of Human IL-2
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The human IL-2 cDNA was subcloned in pQE-30 plasmid and expressed in E. coli M15 cells. Polyclonal antibodies to the N-terminus of Fas (N-18) and C-terminus of FasL (С-178) were from Sigma (United States); CD4-PE, CD4-FITC, CD4-TC, CD25-FITC, CD127-PE, FasL-FITC, FasL-PE, and Fas-PE were from Caltag Medsystems (Buckingham, UK). Control antibodies were IgG from rabbit serum (Sigma-Aldrich, St. Louis, MO), following the methods of Sashchenko et al. [25 (link)].
4
Recombinant Tag7 Purification and Characterization
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The cDNAs encoding recombinant human Tag7 (GenBank accession No. NM_005091) were subcloned in pQE-31 and expressed in E. coli М15 (pREP4; Qiagen, Hilden, Germany). Tag7 was purified as described previously [22] . Limulus assays (Limulus amebocyte lysis chromogenic endpoint assay; Cambrex) of recombinant Tag7 did not detect LPS above the detection limit of 1 endotoxin unit per microgram of protein. In some of the experiments we used Tag7 purified from the human lymphocytes. For this purpose, we gently lysed human lymphocytes via freeze thawing, then the cytosolic fraction was collected in the presence of protease inhibitors, and loaded onto the anti-Tag7 conjugated BrCN-Sepharose. The purity of the eluted protein was assessed by SDS-PAGE. Polyclonal antibodies to the Nterminus of Fas (N-18), C-terminus of FasL (С-178), NKG2D, and MicA were from Sigma-Aldrich (St. Louis, MO, USA); CD4-FITC, CD4-PE, CD8-FITC, CD8-TC, CD3-PE, CD3-TC, CD16-FITC, CD56-PE, NKG2D-FITC, MicA-PE, FasL-FITC, FasL-PE, and Fas-PE antibodies were from Caltag Medsystems (Buckingham, UK).
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