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human colo 320dm whole cell lysate  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology human colo 320dm whole cell lysate
FIGURE 5. Expression of OB-R in the thymus. A, Ob-Rb mRNA ex- pression was examined in the thymus 24 h after injection of saline (control) or 200 g/g hydrocortisone to WT C57BL/6 mice. Left panels, Represen- tative flow cytometric analyses of thymocytes from control (top) or hy- drocortisone (bottom)-injected mice stained for CD4 and CD8. Total thy- mocyte numbers were 45 106 cells (control) and 4 106cells (hydrocortisone). The right panels show Ob-rb (top) and actin (bottom) mRNA expression as assessed by RT-PCR. Lane 1, Control thymus; lane 2, hydrocortisone thymus; lane 3, negative control, PCR performed on non-reverse-transcribed RNA from control thymus. The position of mo- lecular mass markers (bp) is indicated on the right. B, Expression of WT and db alleles of Ob-rb mRNA was examined by RT-PCR on thymus RNA obtained from two db/db to WT BMCs. A schematic representation of the two alleles with the location of the PCR primers (arrows) is shown on the left. The db mutation () leads to insertion of an additional 106-bp exon (f), between exons 17 () and 18b (u) of the mouse lepr gene. This additional exon contains a stop codon causing premature termination of the Ob-rb protein in the db mutant (42). Amplification of the WT and db alleles with the indicated primers yields PCR products of 170 and 276 bp, respec- tively. The right panel shows the expression of mRNA encoding both WT (recipient-derived) and db (donor-derived) alleles in the thymus of two different db/db to WT BMCs (lanes 1 and 2). C, Expression of the Ob-rb protein was examined by Western blotting in thymocytes, whole thymus, a thymic stromal fraction, and in the thymic epithelial cell line MTE-4.14 (upper panel). The membrane was stripped and reblotted with anti--actin Abs (lower panel). The positions of molecular mass standards are indicated on the right. Lane 1, Human COLO <t>320DM</t> cell lysate (positive control); lane 2, total thymocytes; lane 3, whole thymus; lane 4, thymic stromal fraction; and lane 5, MTE- 4.14 total cell lysate.
Human Colo 320dm Whole Cell Lysate, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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FIGURE 5. Expression of OB-R in the thymus. A, Ob-Rb mRNA ex- pression was examined in the thymus 24 h after injection of saline (control) or 200 g/g hydrocortisone to WT C57BL/6 mice. Left panels, Represen- tative flow cytometric analyses of thymocytes from control (top) or hy- drocortisone (bottom)-injected mice stained for CD4 and CD8. Total thy- mocyte numbers were 45 106 cells (control) and 4 106cells (hydrocortisone). The right panels show Ob-rb (top) and actin (bottom) mRNA expression as assessed by RT-PCR. Lane 1, Control thymus; lane 2, hydrocortisone thymus; lane 3, negative control, PCR performed on non-reverse-transcribed RNA from control thymus. The position of mo- lecular mass markers (bp) is indicated on the right. B, Expression of WT and db alleles of Ob-rb mRNA was examined by RT-PCR on thymus RNA obtained from two db/db to WT BMCs. A schematic representation of the two alleles with the location of the PCR primers (arrows) is shown on the left. The db mutation () leads to insertion of an additional 106-bp exon (f), between exons 17 () and 18b (u) of the mouse lepr gene. This additional exon contains a stop codon causing premature termination of the Ob-rb protein in the db mutant (42). Amplification of the WT and db alleles with the indicated primers yields PCR products of 170 and 276 bp, respec- tively. The right panel shows the expression of mRNA encoding both WT (recipient-derived) and db (donor-derived) alleles in the thymus of two different db/db to WT BMCs (lanes 1 and 2). C, Expression of the Ob-rb protein was examined by Western blotting in thymocytes, whole thymus, a thymic stromal fraction, and in the thymic epithelial cell line MTE-4.14 (upper panel). The membrane was stripped and reblotted with anti--actin Abs (lower panel). The positions of molecular mass standards are indicated on the right. Lane 1, Human COLO 320DM cell lysate (positive control); lane 2, total thymocytes; lane 3, whole thymus; lane 4, thymic stromal fraction; and lane 5, MTE- 4.14 total cell lysate.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Indirect effects of leptin receptor deficiency on lymphocyte populations and immune response in db/db mice.

doi: 10.4049/jimmunol.177.5.2899

Figure Lengend Snippet: FIGURE 5. Expression of OB-R in the thymus. A, Ob-Rb mRNA ex- pression was examined in the thymus 24 h after injection of saline (control) or 200 g/g hydrocortisone to WT C57BL/6 mice. Left panels, Represen- tative flow cytometric analyses of thymocytes from control (top) or hy- drocortisone (bottom)-injected mice stained for CD4 and CD8. Total thy- mocyte numbers were 45 106 cells (control) and 4 106cells (hydrocortisone). The right panels show Ob-rb (top) and actin (bottom) mRNA expression as assessed by RT-PCR. Lane 1, Control thymus; lane 2, hydrocortisone thymus; lane 3, negative control, PCR performed on non-reverse-transcribed RNA from control thymus. The position of mo- lecular mass markers (bp) is indicated on the right. B, Expression of WT and db alleles of Ob-rb mRNA was examined by RT-PCR on thymus RNA obtained from two db/db to WT BMCs. A schematic representation of the two alleles with the location of the PCR primers (arrows) is shown on the left. The db mutation () leads to insertion of an additional 106-bp exon (f), between exons 17 () and 18b (u) of the mouse lepr gene. This additional exon contains a stop codon causing premature termination of the Ob-rb protein in the db mutant (42). Amplification of the WT and db alleles with the indicated primers yields PCR products of 170 and 276 bp, respec- tively. The right panel shows the expression of mRNA encoding both WT (recipient-derived) and db (donor-derived) alleles in the thymus of two different db/db to WT BMCs (lanes 1 and 2). C, Expression of the Ob-rb protein was examined by Western blotting in thymocytes, whole thymus, a thymic stromal fraction, and in the thymic epithelial cell line MTE-4.14 (upper panel). The membrane was stripped and reblotted with anti--actin Abs (lower panel). The positions of molecular mass standards are indicated on the right. Lane 1, Human COLO 320DM cell lysate (positive control); lane 2, total thymocytes; lane 3, whole thymus; lane 4, thymic stromal fraction; and lane 5, MTE- 4.14 total cell lysate.

Article Snippet: Human COLO 320DM whole cell lysate (Santa Cruz Biotechnology) was used as a positive control for OB-R expression.

Techniques: Expressing, Injection, Saline, Control, Staining, Reverse Transcription Polymerase Chain Reaction, Negative Control, Reverse Transcription, Mutagenesis, Derivative Assay, Western Blot, Membrane, Positive Control