Anti-Phospho-ASK1 (Thr845) Polyclonal Antibody is a Rabbit antibody targeting Phospho-ASK1 (Thr845). Anti-Phospho-ASK1 (Thr845) Polyclonal Antibody can be used in WB,FCM.

IgG

Human,Mouse,Rat

1. Blank control (blue): Hela (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody: Rabbit Anti-phospho-ASK1 (Thr845) antibody (TMAB-01392), Dilution: 0.2 μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit Igg (orange),used under the same conditions);
Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
2. Sample:
Lane1: Brain (Rat) Lysate at 30 μg
Lane2: Heart (Rat) Lysate at 30 μg
Primary: Anti-phospho-ASK1 (Thr845) (TMAB-01392) at 1:200 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit Igg (secondary antibody) at 1: 3000 dilution;
Predicted band size: 155 kDa
Observed band size: 155 kDa
3. Sample:
Lane 8: Cerebrum (Rat) Lysate at 40 μg
Lane 9: Stomach (Rat) Lysate at 40 μg
Primary: Anti-phospho-ASK1 (Thr845) (TMAB-01392) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 154 kDa
Observed band size: 154 kDa
4. Blank control: U937. Primary Antibody (green line): Rabbit Anti-phospho-ASK1 (Thr845) antibody (TMAB-01392)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Blank control: Mouse spleen.
Primary Antibody (green line): Rabbit Anti-phospho-RelB (Ser551) antibody (TMAB-01392)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB: 1:500-2000; FCM: 1μg /test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human ASK1 around the phosphorylation site of Thr845

Human

MAP3K5

Mitogen-activated protein kinase kinase kinase 5

Apoptosis,Hypertrophy,Kinases

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. Plays an important role in the cascades of cellular responses evoked by changes in the environment. Mediates signaling for determination of cell fate such as differentiation and survival. Plays a crucial role in the apoptosis signal transduction pathway through mitochondria-dependent caspase activation. MAP3K5/ASK1 is required for the innate immune response, which is essential for host defense against a wide range of pathogens. Mediates signal transduction of various stressors like oxidative stress as well as by receptor-mediated inflammatory signals, such as the tumor necrosis factor (TNF) or lipopolysaccharide (LPS). Once activated, acts as an upstream activator of the MKK/JNK signal transduction cascade and the p38 MAPK signal transduction cascade through the phosphorylation and activation of several MAP kinase kinases like MAP2K4/SEK1, MAP2K3/MKK3, MAP2K6/MKK6 and MAP2K7/MKK7. These MAP2Ks in turn activate p38 MAPKs and c-jun N-terminal kinases (JNKs). Both p38 MAPK and JNKs control the transcription factors activator protein-1 (AP-1).