bsm-52189R
[Primary Antibody]
PKR(T446) (10A1) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: PKR(T446)
Modification Site: T446
Clonality: Monoclonal
Isotype: IgG
Entrez Gene: 5610
Swiss Prot: P19525
Source: Synthetic peptide derived from human PKR(T446), around 450-500aa (phospho T446).
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Background:
IFN-induced dsRNA-dependent serine/threonine-protein kinase which plays a key role in the innate immune response to viral infection and is also involved in the regulation of signal transduction, apoptosis, cell proliferation and differentiation. Exerts its antiviral activity on a wide range of DNA and RNA viruses including hepatitis C virus (HCV), hepatitis B virus (HBV), measles virus (MV) and herpes simplex virus 1 (HHV-1). Inhibits viral replication via phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (EIF2S1), this phosphorylation impairs the recycling of EIF2S1 between successive rounds of initiation leading to inhibition of translation which eventually results in shutdown of cellular and viral protein synthesis. Also phosphorylates other substrates including p53/TP53, PPP2R5A, DHX9, ILF3, IRS1 and the HHV-1 viral protein US11. In addition to serine/threonine-protein kinase activity, also has tyrosine-protein kinase activity and phosphorylates CDK1 at 'Tyr-4' upon DNA damage, facilitating its ubiquitination and proteosomal degradation. Either as an adapter protein and/or via its kinase activity, can regulate various signaling pathways (p38 MAP kinase, NF-kappa-B and insulin signaling pathways) and transcription factors (JUN, STAT1, STAT3, IRF1, ATF3) involved in the expression of genes encoding proinflammatory cytokines and IFNs. Activates the NF-kappa-B pathway via interaction with IKBKB and TRAF family of proteins and activates the p38 MAP kinase pathway via interaction with MAP2K6. Can act as both a positive and negative regulator of the insulin signaling pathway (ISP). Negatively regulates ISP by inducing the inhibitory phosphorylation of insulin receptor substrate 1 (IRS1) at 'Ser-312' and positively regulates ISP via phosphorylation of PPP2R5A which activates FOXO1, which in turn up-regulates the expression of insulin receptor substrate 2 (IRS2).
Size: 100ul
Concentration: 1ug/ul
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Lane 1: Hela treated with Calyculin A and TNF-alpha whole cell lysates; Lane 2: Untreated Hela whole cell lysates; probed with PKR(T446) (10A1) Monoclonal Antibody (bsm-52189R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:200 overnight at 4°C, DAB staining.
test
Lane 1: Human Hela cell lysates; Lane 2: Human Hcclm3 cell lysates probed with PKR(T446) (10A1) Monoclonal Antibody, Unconjugated (bsm-52189R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.