- Производитель:
- Sigma-Aldrich
Кат. номер |
ZRB1434-25UL |
ZRB1434-4X25UL |
Specificity
Clone 1B17 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Checkpoint kinase 2 (Chk2) phosphorylated on Threonine 68.
Evaluated by Western Blotting in lysate from HeLa cells treated with UV light.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Checkpoint kinase 2 (Chk2) in lysate from HeLa cells treated with UV light (50 milli Joules followed by recovery of 1 h), but not in untreated cells.
Tested applications
Peptide Inhibition Assay: Target band detection in lysate from UV light treated HeLa cells was prevented by pre-blocking of a representative lot with the phosphorylated Chk2-Thr68 peptide, but not the corresponding non-phosphopeptide.
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-Chk2-Thr68 in human colon cancer tissue sections.
Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Chk2-Thr68 in HeLa cells treated with UV light.
Affinity Binding Assay: A representative lot of this antibody bound phospho-Chk2-Thr68 peptide with a KD of 2.5 x 10-7 in an affinity binding assay.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal, Cat. No. ZRB1434, detects phospho-Chk2-Thr68 and is used in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry, and Peptide Inhibition Assay, and Western Blotting.
Target description
Serine/threonine-protein kinase Chk2 (UniProt: O96017; also known as EC:2.7.11.1, CHK2 checkpoint homolog, Cds1 homolog, Hucds1, hCds1, Checkpoint kinase 2, Chk2) is encoded by the CHEK2 (also known as CDS1, CHK2, RAD53) gene (Gene ID: 11200) in human. Chk2 is a homodimeric nuclear serine/threonine protein kinase that is required for checkpoint-mediated cell cycle arrest, activation of DNA repair, and apoptosis in response to the presence of DNA double-strand breaks. Its protein kinase domain is localized to amino acids 220-486. It is highly expressed in testis, spleen, colon, and peripheral blood leukocytes and lower expression is found in other tissues. Following activation, Chk2 phosphorylates several effector molecules preferentially at the consensus sequence [L-X-R-X-X-S/T]. It is reported to regulate cell cycle checkpoint arrest through phosphorylation of Cdc25A, Cdc25B, and Cdc25C, inhibiting their activity, which can lead to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. Chk2 undergoes phosphorylation at serine 73 by polo-like kinase 3 in response to DNA damage, promoting phosphorylation at threonine 68 by ATM kinase. Phosphorylation at threonine 68 induces its homodimerization and allows autophosphorylation to proceed at threonine 383 and 387 in the T-loop/activation segment that makes it fully active. activation state . Thirteen different isoforms of Chk2 have been described that are produced by alternative splicing. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb® formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Reconstitution
300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Storage and Stability
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Quality Level | 200 |
biological source | rabbit |
recombinant | expressed in HEK 293 cells |
antibody form | purified antibody |
antibody product type | primary antibodies |
clone | 1B17, recombinant monoclonal |
description | recombinant, expressed in HEK 293 cells |
product line | ZooMAb® |
form | lyophilized |
mol wt | calculated mol wt 60.92 kDa","observed mol wt ~60 kDa |
purified by | using Protein A |
species reactivity | human |
species reactivity (predicted by homology) | monkey |
packaging | antibody small pack of 25 µL |
enhanced validation | recombinant expression |
application(s) | affinity binding assay: suitable","immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable","western blot: suitable |
isotype | IgG |
epitope sequence | N-terminal |
conjugate | unconjugated |
Protein ID accession no. | NP_665861, |
UniProt accession no. | O96017, |
shipped in | ambient |
storage temp. | 2-8°C |
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