goat anti guinea pig igg polyclonal cf 488a  (Biotium)

Journal: Skin Health and Disease

Article Title: Pigment epithelium derived factor drives melanocyte proliferation and migration in neurofibromatosis café au lait macules

doi: 10.1002/ski2.394

Figure Lengend Snippet: Study design, genotypes and NF1 expression. (a) Study outline. (b) Somatic mutations within CALMs melanocytes. (c, d) NF1 expression levels within unaffected relative to CALMs melanocytes, 5 replicates per patient sample, relative to normal melanocytes (dotted line). CALMs, café au lait macules; NF1, neurofibromatosis type 1.

Article Snippet: Membranes were blocked with 10% bovine serum albumin (BSA, Sigma‐Aldrich) or 10% skimmed milk and probed using the following antibodies: NF1 (H12, Santa Cruz Biotechnology and D7R7D, Cell Signalling Technology), pERK (Erk1/2, Cell Signalling Technology), total ERK (Erk1/2, Cell Signalling Technology), pAKT (D9E, Cell Signalling Technology), total AKT (C67E7, Cell Signalling Technology), GAPDH (MAB374, Cell Signalling Technology), secondary antibodies (ab98693 and ab97051, Abcam).

Techniques: Expressing

Journal: Skin Health and Disease

Article Title: Pigment epithelium derived factor drives melanocyte proliferation and migration in neurofibromatosis café au lait macules

doi: 10.1002/ski2.394

Figure Lengend Snippet: Comparative transcriptomic analysis of Classic neurofibromatosis 1, 3bp deletion neurofibromatosis 1 and Legius syndrome. (a) Principal component analysis plot of individual patient CALMs transcriptional profiles based on genotype. (b) Heatmap showing Gene Set Enrichment Analysis data using Hallmark gene sets of averaged differential expression genes for each genotype. (c) qRT‐PCR comparing CALMs versus unaffected skin melanocyte mRNA of classic and 3bp deletion neurofibromatosis 1 for individual downstream Ras/mitogen‐activated protein kinase regulated genes shown as averages. CALMs, café au lait macules.

Article Snippet: Membranes were blocked with 10% bovine serum albumin (BSA, Sigma‐Aldrich) or 10% skimmed milk and probed using the following antibodies: NF1 (H12, Santa Cruz Biotechnology and D7R7D, Cell Signalling Technology), pERK (Erk1/2, Cell Signalling Technology), total ERK (Erk1/2, Cell Signalling Technology), pAKT (D9E, Cell Signalling Technology), total AKT (C67E7, Cell Signalling Technology), GAPDH (MAB374, Cell Signalling Technology), secondary antibodies (ab98693 and ab97051, Abcam).

Techniques: Quantitative Proteomics, Quantitative RT-PCR

Journal: Skin Health and Disease

Article Title: Pigment epithelium derived factor drives melanocyte proliferation and migration in neurofibromatosis café au lait macules

doi: 10.1002/ski2.394

Figure Lengend Snippet: Café au Lait Macules characterised by decreased expression of pigment epithelium‐derived factor. (a) Venn diagram of unfiltered differentially expressed genes shared between the two NF1 genotypes. (b) qRT‐PCR for the PEDF gene SERPINF1 comparing CALMs mRNA of classic and 3bp deletion neurofibromatosis 1 versus normal melanocytes. (c) qRT‐PCR for the PEDF gene, SERPINF1 , expression comparing normal melanocytes treated with NF1‐targeting and non‐targeting siRNA. (d) Migration assay comparing normal melanocytes and those treated with NF1‐targeting and non‐targeting siRNA. NF1, neurofibromatosis type 1; PEDF, pigment epithelium‐derived factor.

Article Snippet: Membranes were blocked with 10% bovine serum albumin (BSA, Sigma‐Aldrich) or 10% skimmed milk and probed using the following antibodies: NF1 (H12, Santa Cruz Biotechnology and D7R7D, Cell Signalling Technology), pERK (Erk1/2, Cell Signalling Technology), total ERK (Erk1/2, Cell Signalling Technology), pAKT (D9E, Cell Signalling Technology), total AKT (C67E7, Cell Signalling Technology), GAPDH (MAB374, Cell Signalling Technology), secondary antibodies (ab98693 and ab97051, Abcam).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Migration

Journal: Skin Health and Disease

Article Title: Pigment epithelium derived factor drives melanocyte proliferation and migration in neurofibromatosis café au lait macules

doi: 10.1002/ski2.394

Figure Lengend Snippet: PEDF rescues loss of NF1 associated proliferation and migration. (a) Characterisation of MM cell lines. qRT‐PCR (b) and Western blot analysis (c) for NF1 expression in MM cell lines. (d) qRT‐PCR for MITF and PEDF gene expression in MM cell lines. (e) Validation of NF1‐targeting siRNA treatment of M14 melanoma cells by qRT‐PCR. Functional assessments of M14 melanoma cell proliferation (f) and invasion (g) after NF1 knockdown and PEDF rescue. MM, malignant melanoma; NF1, neurofibromatosis type 1; PEDF, pigment epithelium‐derived factor.

Article Snippet: Membranes were blocked with 10% bovine serum albumin (BSA, Sigma‐Aldrich) or 10% skimmed milk and probed using the following antibodies: NF1 (H12, Santa Cruz Biotechnology and D7R7D, Cell Signalling Technology), pERK (Erk1/2, Cell Signalling Technology), total ERK (Erk1/2, Cell Signalling Technology), pAKT (D9E, Cell Signalling Technology), total AKT (C67E7, Cell Signalling Technology), GAPDH (MAB374, Cell Signalling Technology), secondary antibodies (ab98693 and ab97051, Abcam).

Techniques: Migration, Quantitative RT-PCR, Western Blot, Expressing, Gene Expression, Biomarker Discovery, Functional Assay, Knockdown, Derivative Assay