- Производитель:
- Sigma-Aldrich
Кат. номер |
ZRB2028-4X25UL |
ZRB2028-25UL |
Specificity
Clone 4K6 is a Rabbit recombinant monoclonal antibody that detects all three isoforms of AKT. It targets an epitope within 20 amino acids from the C-terminal region.
Quality Control Testing
Evaluated by Western Blotting in MCF-7 cell lysate.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected AKT1 in MCF-7 cell lysate.
Tested applications
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected AKT1 in NIH3T3 and C6 cell lysates.
Affinity Binding Assay: A representative lot of this antibody bound pan AKT1 peptide with a KD of 1.0 x 10-12 in an affinity binding assay.
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected AKT1 in human prostate cancer tissue sections.
Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected AKT1 in HeLa, A431, and HepG2 cells.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Target description
Protein Kinase B a (UniProt: P31749; also known as Akt1) are encoded by the AKT1 gene (Gene IDs: 207) in human. Akt, a serine/threonine kinase, is a critical enzyme in several signal transduction pathways involved in cell proliferation, apoptosis, angiogenesis, and diabetes. In mammals three isoforms of Akt have been reported (Akt 1, 2, 3) that exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. Akt1 plays an important role in growth and anti-apoptosis; Akt2 acts primarily as a regulator of glucose metabolism. Although Akt3 does not contribute significantly to the maintenance of normal metabolism in tissues but is essential for the attainment of normal organ size, probably influencing both cell size and number via mTOR activation. Each Akt isoform is composed of three functionally distinct regions: An N-terminal pleckstrin homology (PH) domain that provides a lipid-binding module to direct Akt to PIP2 and PIP3, a central catalytic domain, and a C-terminal hydrophobic motif. Activation of Akt involves growth factor binding to a receptor tyrosine kinase and activation of PI 3-kinase that phosphorylates membrane bound PIP2 to generate PIP3. The binding of PIP3 to the PH domain anchors Akt to the plasma membrane. Akt is activated following its phosphorylation at Thr308 on the kinase domain by PDK1 and on Ser473 on the hydrophobic motif by mTORC2. The principal role of Akt is to facilitate growth factor-mediated cell survival and to block apoptotic cell death. Akt achieves this by phosphorylating a variety of substrates, such as Bad, caspase-9, Forkhead transcription factors, and GSK-3. 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 | 4K6, recombinant monoclonal |
description | recombinant, expressed in HEK 293 cells |
product line | ZooMAb® |
form | lyophilized |
mol wt | calculated mol wt 55.69 kDa","observed mol wt ~56 kDa |
purified by | using Protein A |
species reactivity | mouse, rat, human |
packaging | antibody small pack of 25 µL |
enhanced validation | recombinant expression |
application(s) | affinity binding assay: suitable","immunocytochemistry: suitable","immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable","western blot: suitable |
isotype | IgG |
epitope sequence | C-terminal |
conjugate | unconjugated |
Protein ID accession no. | NP_005154.2, |
UniProt accession no. | P31749, |
shipped in | ambient |
storage temp. | 2-8°C |
WGK Germany | WGK 2 |
Flash Point F | Not applicable |
Flash Point C | Not applicable |
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