Anti-Akt Antibody, PH Domain Antibody, clone SKB1 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1027 - recombinant, expressed in HEK 293 cells
Synonym: EC:2.7.11.1; PKB; PKB alpha; Protein kinase B; Protein kinase B alpha; Proto-oncogene c-Akt; RAC-PK-alpha; RAC-alpha serine/
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was observed in neurons and neuropil of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse and HRP-DAB. Cytoplasmic/membranous staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cell line was performed using a 1:100 dilution of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm and nucleus.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from U87-MG (Lane 1), NIH3T3 (Lane 2), and L6 (Lane 3) cells were probed with Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal (1:1,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Akt (~55 kDa).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million A431 Cells was performed using 1 μg of Cat. No. ZMS1027, Anti-Akt, PH Domain, clone SKB1 ZooMAb® Mouse Monoclonal (Yellow histogram) or the equivalent amount of a Mouse IgG1 isotype control (Grey histogram), followed by a PE-conjugated Anti-Mouse secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | recombinant monoclonal |
| SKB1, monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG1 |
| mol wt | calculated mol wt 55.69 kDa |
| observed mol wt ~55 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, human, mouse |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P31749  |
| Application: | Anti-Akt, PH Domain, clone SKB1 ZooMAb, Cat. No. ZMS1027, is a recombinant Mouse monoclonal antibody that specifically targets Akt and is tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), Flow Cytometry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Akt in U87-MG and L6 cell lysates. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Akt in HeLa and NIH 3T3 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Akt in human brain and human testis tissue sections. Flow Cytometry Analysis: 1 μg from a representative lot detected Akt in 1X10E6 A431 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | GST-tagged recombinant fragment corresponding to the first 149 amino acids from the N-terminal region of human Akt. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. 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. |
| Specificity: | Clone SKB1 is a ZooMAb mouse recombinant monoclonal antibody that specifically detects Akt. It targets an epitope within the N-terminal domain. It displays highest reactivity with Akt1 and lower with Akt2 and Akt3. |
| 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. |
| Target description: | RAC-alpha serine/threonine-protein kinase (UniProt: P31749; also known as EC:2.7.11.1, Protein kinase B, PKB, Protein kinase B alpha, PKB alpha, Proto-oncogene c-Akt, RAC-PK-alpha, Akt) is encoded by the AKT1 (also known as PKB, RAC) gene (Gene ID: 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. |
| WGK Germany | WGK 1 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |