Journal:

Article Title: Salicylhydroxamic Acid Functionalized Affinity Membranes for Specific Immobilization of Proteins and Oligonucleotides

doi:

Figure Lengend Snippet: Release of PDBA-modified protein immobilized on SHA-affinity membranes. Versalinx Protein Releasing Reagent (100 μL) was added to spin columns containing immobilized PDBA-AP, and AP and the columns were incubated at 37°C for 30 min. The releasing reagent was passed through the membranes by slow centrifugation. AP activity in the flow-through was analyzed using pNPP and monitoring absorbance at l = 405 nm. Protein concentrations were determined by preparing standard curves of AP at 0 μg/mL, 10 μg/mL, 20 μg/mL, 30 μg/mL and 40 μg/mL.

Article Snippet: PDBA-human IgG or unmodified human IgG (50 μg/mL) were spotted in Versalinx Spotting Solution (Prolinx) in a 4 × 30 microarray format, 300 pL/spot, 500 μm pitch, using a Packard BioChip Arrayer (PerkinElmer Life Sciences) according to manufacturer’s specifications.

Techniques: Modification, Incubation, Centrifugation, Activity Assay

Journal:

Article Title: Salicylhydroxamic Acid Functionalized Affinity Membranes for Specific Immobilization of Proteins and Oligonucleotides

doi:

Figure Lengend Snippet: Immobilization of PBA-modified oligonucleotides on SHA-affinity membranes. Radiolabeled PBA-modified or -unmodified oligonucleotides (1000 nM in 30 μL 10× SSC) were bound to SHA-affinity membranes for 15 min at room temperature. After binding, membranes were washed, and bound oligonucleotides were released by adding 60 μL 1 mM Versalinx Nucleic Acid Releasing Reagent in distilled water and incubating at 37°C for 10 min. Bound and released oligonucleotides were determined by scintillation counting of membrane, flow-through, and input fractions, and are expressed as a percentage of input fractions.

Article Snippet: PDBA-human IgG or unmodified human IgG (50 μg/mL) were spotted in Versalinx Spotting Solution (Prolinx) in a 4 × 30 microarray format, 300 pL/spot, 500 μm pitch, using a Packard BioChip Arrayer (PerkinElmer Life Sciences) according to manufacturer’s specifications.

Techniques: Modification, Binding Assay, Membrane

Journal:

Article Title: Salicylhydroxamic Acid Functionalized Affinity Membranes for Specific Immobilization of Proteins and Oligonucleotides

doi:

Figure Lengend Snippet: SHA-affinity membranes as microarray surfaces. PDBA-human IgG or unmodified human IgG (50 μg/mL) were spotted onto SHA-affinity membranes in a 4 × 30 microarray format using standard arrayer protocol. The membrane was incubated at room temperature for 1 h, washed with Versalinx Wash Buffer #1, and then developed with 0.1 μg/mL Cy3-goat anti-human IgG for 30 min at room temperature. The array was visualized on a ScanArray Lite at a laser power of 75%, PMT gain of 65%, and spatial resolution of 10 μm.

Article Snippet: PDBA-human IgG or unmodified human IgG (50 μg/mL) were spotted in Versalinx Spotting Solution (Prolinx) in a 4 × 30 microarray format, 300 pL/spot, 500 μm pitch, using a Packard BioChip Arrayer (PerkinElmer Life Sciences) according to manufacturer’s specifications.

Techniques: Microarray, Membrane, Incubation

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: HDM4 (HDMX) is widely expressed in adult pre-B acute lymphoblastic leukemia and is a potential therapeutic target.

doi: 10.1038/modpathol.3800727

Figure Lengend Snippet: Figure 1 Immunohistochemical analysis of HDM4 in bone marrow biopsy specimens from pre-B ALL patients. (a) Hematoxylin and eosin (H&E) stain ( 1000); (b–e) immunostains for p53 (b), HDM2 (c), HDM4 (d), and p21 (e) ( 200).

Article Snippet: Two representative 0.6-mm cores were then obtained from each sample and inserted in a grid pattern into a recipient paraffin block by using a tissue arrayer (Beecher Instruments, Silver Spring, MD, USA) as described elsewhere.18 We stained sections from each tissue microarray using immunohistochemical methods described previously.18 The antibodies used were specific for p53 (1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p21 (1:75; NeoMarkers, Fremont, CA, USA), HDM2 (1:75; Santa Cruz Biotechnology), and HDM4 (1:200; rabbit polyclonal against human recombinant glutathione-S-transferase (GST)MDMXD109-198 protein antibody generated by Dr Guillermina Lozano, manuscript submitted).

Techniques: Immunohistochemical staining, Staining