Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin]

ELISA: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] [NB720-B] - ELISA results of purified Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] tested against purified Mouse IgG. Each well was coated in duplicate with 1.0 ug of Mouse IgG (green line). The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using blocking buffer, Streptavidin HRP conjugate 1:10000, and TMB substrate.

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] [NB720-B] -

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] [NB720-B] - (A) Confocal microscopy of NCTC liver cells before & after exposure to amifostine (100 μg/ml), for double PDH(red)/phosphoPDH(green), phosphoPDH(red), LDH5 (red) & HIF1 alpha (red) expression. Western blot bands following exposure to 100 & 500 μg/ml of amifostine at 30 min is also shown. (B) Western blot expression of phosphoPDH, PDK1, LDH5 & HIF1 alpha in hepatoma HepG2 cells, at 0, 30 & 60 min following exposure to 100 μg/ml of amifostine. (C) Confocal microscopy of NCTC liver cells before & after exposure to amifostine (100 μg/ml), for HIF1 alpha (green), LDH5 (red), PDK1 (green) & phosphoPDH(red) expression with & without silencing of the HIF1 alpha gene. (D) Acetyl-CoA levels (pmol) in NCTC cells at 0, 30 & 60 min following exposure to 100 μg/ml of amifostine. (E) ATP levels (pmol) in NCTC cells at 0, 30 & 60 min following exposure to 100 μg/ml of amifostine. (F) Time course recording of NCTC & HepG2 cell mitochondrial membrane potentials as assessed with the JC1 method & confocal imaging (0–20 minutes), showed a rapid transient reduction of green (monomer) & red (aggregate) forms of the dye that was subsequently restored to normal levels. (G) Mitochondrial ROS (mtROS) production by NCTC & HepG2 cells, after exposure to 18Gy of ionizing radiation with & without pre-incubation with amifostine, showing a strong effect of amifostine in normal NCTC cells. mtROS were low in hepatoma HepG2 cells compared to NCTC hepatocytes & were increased only in dividing neoplastic cells. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27507219), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] [NB720-B] -

Western Blot: Rabbit anti-Mouse IgG (H+L) Secondary Antibody [Biotin] [NB720-B] - (A) Western blot images & band densitometry analysis of levels of proteins involved in anaerobic metabolism, as assessed in mouse liver before & after administration of amifostine. Bars show standard deviation & asterisks refer to p-values (*p < 0.05, **p < 0.001). (B) Confocal immunofluorescent microscopy after staining with anti-PDK1/red antibody showing increased cytoplasmic expression in mouse hepatocytes from 0 (i) to 30 min (ii) & regression thereafter (iii). Double immunostaining with PDH/red & phosphorylated pPDH/green (iv), showed an intensification of the expression of the inactive pPDH form of the enzyme 30 min (v) following amifostine injection & trend for restoration of normal PDH levels at 60 min (vi). Confocal immunofluorescent microscopy after staining for LDH1/red showed stable levels of expression in mouse hepatocytes (i,ii,iii). In contrast, LDH5/red expression was sharply induced 30 min following amifostine injection & decreased thereafter (iv, v, vi). Similar patterns were noted for GLUT2 expression (i,ii,iii) & for HIF1 alpha expression (iv,v,iv). (C) Analysis of the fluorescence intensity of confocal microscopy images (from five representative tissue areas for each staining). Bars show standard deviation & asterisks refer to p-values (*p < 0.05, **p < 0.001). (D) mRNA expression levels of LDHA, PDK1 & GLUT2 (three mice for each time point) following exposure to amifostine, as measured with quantitative RT-PCR. Bars show standard deviation & asterisks refer to p-values (**p < 0.001). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27507219), licensed under a CC-BY license. Not internally tested by Novus Biologicals.