multiple human adult brain tissue northern blots  (TaKaRa)

Journal: Breast Cancer Research

Article Title: Involvement of maternal embryonic leucine zipper kinase (MELK) in mammary carcinogenesis through interaction with Bcl-G, a pro-apoptotic member of the Bcl-2 family

doi: 10.1186/bcr1650

Figure Lengend Snippet: Expression and distribution of MELK in human normal tissues and breast cancer cell lines. (a) Expression of MELK in 12 breast cancer specimens (case number; 42, 102, 247, 252, 302, 473, 478, 502, 552, 646, 769 and 779) by semi-quantitative RT-PCR. GAPDH served as a quantitative internal control. (b) Multiple tissue Northern blot analysis demonstrated that an approximately 2.7 kb MELK transcript was detected in the testis, thymus and small intestine. PBL, peripheral blood leukocytes. (c) Breast cancer cell line Northern blot analysis revealed that approximately 2.4 to 2.7 kb MELK variants were specifically expressed in breast cancer cell lines, but not in normal vital organs. (d) Schematic representation of three variant transcripts identified by cDNA library screening (see Materials and methods). White boxes indicate a coding region and black boxes indicate a non-coding region. Black and grey triangles indicate initiation codons, and white triangles indicate stop codons. Exon numbers are shown above each box. (e) In vitro translation assay of each variant isolated from cDNA library screening. The number within parentheses represents the predicted molecular weight (kDa) of each variant protein. (f) Expression of MELK proteins in eight breast cancer cell lines as well as human mammary epithelial cells (HMECs) shown by western blot analysis with an anti-MELK antibody. β-Actin served as a control. (g) Schematic representation of the V1, V2 and V3 forms of MELK. The shaded boxes indicate the catalytic domain (amino acids 11 to 263 of the V1 protein). The KA1 domain is the kinase-associated domain in the carboxy-terminal region.

Article Snippet: Breast cancer Northern blots and human multiple-tissue Northern blots (Takara Clontech) were hybridized with [α 32 P]-dCTP-labeled PCR products of MELK cDNA prepared by RT-PCR (see below).

Techniques: Expressing, Quantitative RT-PCR, Northern Blot, Variant Assay, cDNA Library Assay, In Vitro, Isolation, Molecular Weight, Western Blot

Journal: Genomics Insights

Article Title: Cloning and Initial Functional Characterization of Mlk4α and Mlk4β

doi: 10.4137/GEI.S6092

Figure Lengend Snippet: Expression analysis of MLK4α and MLK4β : A ) Northern blot hybridization for MLK4β , B ) PCR with multiple tissue cDNA panel for MLK4α and MLK4β .

Article Snippet: To determine the distribution of MLK4α and MLK4β mRNA within the tissues, a multiple human tissue Northern blot (Clontech, Palo Alto, CA, USA) was hybridized to the non-coding 3′-end of specific DNA fragments of MLK4α (1970–3910 bp) and MLK4β (2087–4667 bp).

Techniques: Expressing, Northern Blot, Hybridization

Journal: BMC Genomics

Article Title: Gene organization, evolution and expression of the microtubule-associated protein ASAP (MAP9)

doi: 10.1186/1471-2164-9-406

Figure Lengend Snippet: Expression of ASAP in human and murine tissues . (A) Northern-blot analysis of ASAP transcript in adult human tissues was carried out with pre-made Northern blots purchased from Clontech. The blots were hybridized with a 32 P-labelled full-length ASAP cDNA probe. The sizes of molecular weight marker are shown on the left. (B) RT-PCR analysis of ASAP in murine tissues. Total RNA was prepared from various mouse tissues. A 279-bp ASAP RT-PCR product is visualized. Expression of GAPD was determined as a control of RNA integrity. (C) Western-blot analysis of mASAP protein was carried out using the mASAP antibody on testis, brain, spleen and colon mouse tissue extracts and compared to the profile observed on NIH3T3 and U2-OS (positive controls) or HT29 (negative control derived from colon) cell extracts. α-tubulin levels served as loading control.

Article Snippet: Two human tissue northern blots (MTN I and MTN II, Clontech) were probed with a random-primed [32P]dCTP-labelled ASAP cDNA.

Techniques: Expressing, Northern Blot, Molecular Weight, Marker, Reverse Transcription Polymerase Chain Reaction, Western Blot, Negative Control, Derivative Assay

Journal: PLoS ONE

Article Title: Upregulation of Nuclear Factor-Related Kappa B Suggests a Disorder of Transcriptional Regulation in Minimal Change Nephrotic Syndrome

doi: 10.1371/journal.pone.0030523

Figure Lengend Snippet: A, Western blot of total lysates using anti-NFRKB reveals a double band at the expected size. The specificity was attested by the loss of the binding upon preincubation of antibody with synthetic peptides corresponding to positions 543–557 and 1091–1106 within the primary structure of NFRKB. These data are representative of three independent experiments. B, Northern blot analysis of NFRKB. Multiple tissue-Northern blot (Clontech Laboratories, Inc.) was hybridized with a full-length-probe and exposed to Phosphoimager Storm for 24 h. This experiment was repeated once.

Article Snippet: Human multiple-tissue Northern blot (CLONTECH Laboratories, Inc.) was hybridized with a cDNA probe corresponding to the full-length coding sequence.

Techniques: Western Blot, Binding Assay, Northern Blot