polyclonal rabbit anti human jnk antibody  (Cell Signaling Technology Inc)

Journal: PLoS ONE

Article Title: PknE, a Serine/Threonine Protein Kinase of Mycobacterium tuberculosis Initiates Survival Crosstalk That Also Impacts HIV Coinfection

doi: 10.1371/journal.pone.0083541

Figure Lengend Snippet: Controls (Ctrl, LPS) and infected cells were lysed post infection at varied time points and subjected to western blotting. The blots were probed with phospho A) Erk1/2, E) p38, and I) SAPK/JNK and their respective non phospho (C, G and I) antibodies. The results are from three independent experiments. Figures B, D, F, H, J, K, and L depict the corresponding densitometry values of phospho and nonphospho antibody probed blots. *, *** denotes p<0.05 and p<0.0001, when Δ pknE infected macrophages compared with Rv (one – way ANOVA). The abbreviations ctrl denotes control and LPS denotes lipopolysaccharide.

Article Snippet: Cell lysates were prepared as reported earlier and the immunoblots were probed with rabbit anti-human polyclonal antibodies (Cell Signaling Technologies) against phospho and non-phospho Akt, p38, Erk½ and SAPK/JNK (1∶1000) and detected using horseradish peroxidase-conjugated goat anti-rabbit antibody (1∶300) (Amersham Biosciences).

Techniques: Infection, Western Blot

Journal: PLoS ONE

Article Title: PknE, a Serine/Threonine Protein Kinase of Mycobacterium tuberculosis Initiates Survival Crosstalk That Also Impacts HIV Coinfection

doi: 10.1371/journal.pone.0083541

Figure Lengend Snippet: Cells to be infected were treated with A) SAPK/JNK and B) p38 inhibitors, for 1 h and lyzed 1 h post infection. The lysates were subjected to western blotting and probed with phospho and non phospho Erk½ antibodies. The results from three independent experiments are shown. The abbreviations Ctrl denote control and LPS denotes lipopolysaccharide.

Article Snippet: Cell lysates were prepared as reported earlier and the immunoblots were probed with rabbit anti-human polyclonal antibodies (Cell Signaling Technologies) against phospho and non-phospho Akt, p38, Erk½ and SAPK/JNK (1∶1000) and detected using horseradish peroxidase-conjugated goat anti-rabbit antibody (1∶300) (Amersham Biosciences).

Techniques: Infection, Western Blot

Journal: PLoS ONE

Article Title: PknE, a Serine/Threonine Protein Kinase of Mycobacterium tuberculosis Initiates Survival Crosstalk That Also Impacts HIV Coinfection

doi: 10.1371/journal.pone.0083541

Figure Lengend Snippet: Controls (ctrl, LPS) and infected cells were lysed post NO stress and subjected to western blotting. The blots were probed with phospho A) Erk1/2, B) p38, and C) SAPK/JNK with their respective non phospho antibodies and the results of three independent experiments are given. The corresponding densitometry values of phospho and nonphospho are given at the end of antibody probed blots. *** denotes p<0.0001, when Δ pknE infected macrophages compared with Rv (one – way ANOVA). The abbreviations ctrl denotes control, LPS denotes lipopolysaccharide, and NO denotes nitric oxide. Nuclear fractions isolated post infection was subjected to DNA binding ELISA D) ATF-2 and E) NF-kB. The results are from three independent experiments. The error bars denote standard error of the means. * denotes p<0.05 (one way ANOVA) when Δ pknE +NO was compared to Rv+NO treated macrophages.

Article Snippet: Cell lysates were prepared as reported earlier and the immunoblots were probed with rabbit anti-human polyclonal antibodies (Cell Signaling Technologies) against phospho and non-phospho Akt, p38, Erk½ and SAPK/JNK (1∶1000) and detected using horseradish peroxidase-conjugated goat anti-rabbit antibody (1∶300) (Amersham Biosciences).

Techniques: Infection, Western Blot, Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: PknE, a Serine/Threonine Protein Kinase of Mycobacterium tuberculosis Initiates Survival Crosstalk That Also Impacts HIV Coinfection

doi: 10.1371/journal.pone.0083541

Figure Lengend Snippet: Human monocyte derived macrophages (n = 6), were infected with M. tuberculosis strains followed by coinfection with a CCR5 tropic virus in the presence (A), and absence of SAPK/JNK inhibitor (B). Similarly, coinfection was performed using CXCR4 tropic virus in the presence (C) and absence of SAPK/JNK inhibitor (D). p24 antigen levels were estimated using ELISA on day 4. * denotes p<0.05 (one way – Anova) when Δ pknE was compared to Rv infected macrophages.

Article Snippet: Cell lysates were prepared as reported earlier and the immunoblots were probed with rabbit anti-human polyclonal antibodies (Cell Signaling Technologies) against phospho and non-phospho Akt, p38, Erk½ and SAPK/JNK (1∶1000) and detected using horseradish peroxidase-conjugated goat anti-rabbit antibody (1∶300) (Amersham Biosciences).

Techniques: Derivative Assay, Infection, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Cross-Linking Mast Cell Specific Gangliosides Stimulates the Release of Newly Formed Lipid Mediators and Newly Synthesized Cytokines

doi: 10.1155/2016/9160540

Figure Lengend Snippet: Cross-linking GD1b derived gangliosides with mAbAA4 induced MAP kinase phosphorylation in mast cells. RBL-2H3 cells were either stimulated via Fc ε RI, where cells were sensitized with IgE anti-TNP and stimulated with DNP 48 -HSA (50 ng/mL) or incubated with mAbAA4 (1, 2.5, 5, and 10 μ g/mL) for 10 min. Total cell lysates were immunoblotted with antibodies against phospho-ERK1/2 (p-ERK1/2), ERK1/2, phospho-JNK1/2 (p-JNK1/2), JNK1/2, phospho-p38 (p-p38), p38, and α / β -tubulin (housekeeping protein) and the mean optical density of the bands was determined. Densitometry of the changes in expression and phosphorylation of proteins were corrected for α / β -tubulin. Data is expressed as the fold of nonstimulated (NS) cells. (a) Ratio of phosphorylated ERK1/2/total ERK1/2; (b) a representative blot from (a); (c) ratio of phosphorylated JNK1/2/total JNK1/2; (d) a representative blot from (c); (e) ratio of phosphorylated p38/total p38; (f) a representative blot from (e). Data is expressed as the mean ± SD of three independent experiments. ∗ P < 0.05 between experimental samples and the nonstimulated (NS) cells. # P < 0.05 between experimental samples and Fc ε RI stimulated cells.

Article Snippet: Rabbit polyclonal antibody anti-human phospho-cPLA 2 ; rabbit polyclonal antibody anti-human cPLA 2 ; rabbit mAb anti-human phospho-ERK1/2; rabbit mAb anti-rat ERK1/2; rabbit mAb anti-human phospho-JNK1/2; rabbit polyclonal antibody anti-human JNK1/2; rabbit mAb anti-human phospho-p38; rabbit polyclonal antibody anti-human anti-p38, and rabbit polyclonal antibody anti-human α / β -tubulin were purchased from Cell Signaling Technology Inc. (Danvers, MA).

Techniques: Derivative Assay, Incubation, Expressing