Product Info Summary
SKU: | MP00024 |
---|---|
Size: | 100ug |
Reactive Species: | Human, Monkey, Mouse, Rat |
Host: | Mouse |
Application: | ELISA, IF, IHC, WB |
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Product info
Product Name
Anti-Akt phospho S473 Monoclonal Antibody
SKU/Catalog Number
MP00024
Size
100ug
Form
Liquid (sterile filtered)
Description
Boster Bio Anti-Akt phospho S473 Monoclonal Antibody (Catalog # MP00024). Tested in ELISA, IF, IHC, WB applications. This antibody reacts with Human, Mouse, Rat, Monkey.
Storage & Handling
Store Anti-AKT pS473 (MOUSE) Monoclonal Antibody at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.
Cite This Product
Anti-Akt phospho S473 Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # MP00024)
Host
Mouse
Contents
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
Clonality
Monoclonal
Clone Number
Clone: 17F6.B11
Isotype
IgG1 kappa
Immunogen
Anti-AKT pS473 (MOUSE) Monoclonal Antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein, followed by hybridoma development.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
MP00024 is reactive to AKT1 in Human, Monkey, Mouse, Rat
Applications
MP00024 is guaranteed for ELISA, IF, IHC, WB Boster Guarantee
Observed Molecular Weight
Calculated molecular weight
55.686kDa
Background of Akt1
AKT phospho S473 is a component of the PI-3 kinase pathway and is activated by phosphorylation at Ser 473 and Thr 308. AKT is a cytoplasmic protein also known as AKT1, Protein Kinase B (PKB) and rac (related to A and C kinases). AKT is a key regulator of many signal transduction pathways. AKT Exhibits tight control over cell proliferation and cell viability. Overexpression or inappropriate activation of AKT is noted in many types of cancer. AKT mediates many of the downstream events of PI 3-kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI 3-kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis; (ii) promotion of proliferation. Anti-AKT pS473 (MOUSE) Monoclonal Antibody is ideal for investigators involved in Cell Signaling, Cancer, Neuroscience, Signal Transduction research.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Innovating Scientists Reward
If you are the first to review this product, or if you have results for a special sample, species or application this product is not validated in, share your results with us and receive product credits you can use towards any Boster products! Applicable to all scientists worldwide.
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Assay dilution & Images
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
ELISA: 1:20,000
Flow Cytometry: User optimized
IHC: 20 µg/mL
IF Microscopy: 1:500-1:3,000
IP: User optimized
WB: 1:500-1:3,000
Validation Images & Assay Conditions
Click image to see more details
Figure 1. IHC analysis of AKT1 using anti-AKT1 antibody (MP00024).
AKT1 was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-AKT1 Antibody (MP00024) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Immunofluorescence confocal microscopy of Mouse Anti-AKT pS473 antibody.Tissue: EGF treated A431 cells.Fixation:0.5% PFA. Antigen retrieval:EGF 15 min.Primary antibody:AKT pS473 antibody at 10 µg/mL
Click image to see more details
High resolution STED immunofluorescence nanoscopy of Mouse anti-AKT pS473 antibody.Tissue:A43the cell.Staining:AKT pS473as red signal with bis-benzimide (blue) nuclear counterstain.
Click image to see more details
Immunofluorescence Microscopy of Mouse Anti-AKTpS473 antibody using STED nanoscopy to evaluate AKT activation and migration.Tissue:A43cells with EGF, a rapid activation of AKT is observed (Panel B) along with a coincident change in the tubulin organization (yellow signal), as well as an extensive cell shape-change (cell membrane folding) and accumulation of AKTpS473 at the cell periphery.
Click image to see more details
Figure 5. Western blot analysis of AKT1 using anti-AKT1 antibody (MP00024).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AKT1 antigen affinity purified polyclonal antibody (Catalog # MP00024) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for AKT1.
Click image to see more details
Figure 6. Western blot analysis of AKT1 using anti-AKT1 antibody (MP00024).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AKT1 antigen affinity purified polyclonal antibody (Catalog # MP00024) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for AKT1.
Click image to see more details
Figure 7. Western blot analysis of AKT1 using anti-AKT1 antibody (MP00024).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AKT1 antigen affinity purified polyclonal antibody (Catalog # MP00024) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for AKT1.
Click image to see more details
Figure 8. Western blot analysis of AKT1 using anti-AKT1 antibody (MP00024).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AKT1 antigen affinity purified polyclonal antibody (Catalog # MP00024) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for AKT1.
Protein Target Info & Infographic
Gene/Protein Information For AKT1 (Source: Uniprot.org, NCBI)
Gene Name
AKT1
Full Name
RAC-alpha serine/threonine-protein kinase
Weight
55.686kDa
Superfamily
protein kinase superfamily
Alternative Names
AKT serine/threonine kinase 1; AKT; Akt1; AKT1m; CWS6; EC 2.7.11; EC 2.7.11.1; PKB alpha; PKB; PKBMGC99656; PRKBA; Protein Kinase B Alpha; Protein kinase B; Proto-oncogene c-Akt; rac protein kinase alpha; RAC; RAC-alpha serine/threonine-protein kinase; RAC-alpha; RAC-PK-alpha; RACPKB-ALPHA; Serine-Threonine Protein Kinase ; v-akt murine thymoma viral oncogene homolog 1; V-Akt Murine Thymoma Viral Oncogene-Like Protein 1 AKT1 AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA AKT serine/threonine kinase 1 RAC-alpha serine/threonine-protein kinase|AKT1m|PKB alpha|RAC-PK-alpha|protein kinase B alpha|proto-oncogene c-Akt|rac protein kinase alpha|serine-threonine protein kinase|v-akt murine thymoma viral oncogene homolog 1|v-akt murine thymoma viral oncogene-like protein 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on AKT1, check out the AKT1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for AKT1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Akt phospho S473 Monoclonal Antibody (MP00024)
Hello CJ!
MP00024 has been cited in 10 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Insulin-like growth factor 2 mRNA-binding protein 1 promotes cell proliferation via activation of AKT and is directly targeted by microRNA-494 in pancreatic cancer
A small interfering RNA targeting osteopontin as gastric cancer therapeutics
Short hairpin RNA targeting AKT1 and PI3K/p85 suppresses the proliferation and self-renewal of lung cancer stem cells
Protective Effects of Sodium (?)-5-Bromo-2-(%u03B1-Hydroxypentyl) Benzoate in a Rodent Model of Global Cerebral Ischemia
Vibration exercise decreases insulin resistance and modulates the insulin signaling pathway in a type 2 diabetic rat model
Epigallocatechin-3-Gallate Promotes the Growth of Mink Hair Follicles Through Sonic Hedgehog and Protein Kinase B Signaling Pathways
Hyperthermia induced HIF-1a expression of lung cancer through AKT and ERK signaling pathways
Epicatechin Plus Treadmill Exercise are Neuroprotective Against Moderate-stage Amyloid Precursor Protein/Presenilin 1 Mice
Progesterone alleviates hypoxic-ischemic brain injury via the Akt/GSK-3? signaling pathway
Li R, Cui K, Liu K, Li H, Zhang Y, Liu X, Chen R, Li M, Wang T, Wang S, Liu J, Rao K. Sci Rep. 2017 Oct 18;7(1):13464. doi: 10.1038/s41598-017-12907-1. Metabolic syndrome in rats is associated with erectile dysfunction by impairing PI3K/Akt/eNOS a...
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Customer Q&As
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15 Customer Q&As for Anti-Akt phospho S473 Monoclonal Antibody
Question
Does anti-Akt phospho S473 Monoclonal antibody MP00024 work for IHC with foreskin?
Verified Customer
Verified customer
Asked: 2020-04-30
Answer
According to the expression profile of foreskin, AKT1 is highly expressed in foreskin. So, it is likely that anti-Akt phospho S473 Monoclonal antibody MP00024 will work for IHC with foreskin.
Boster Scientific Support
Answered: 2020-04-30
Question
I was wanting to use your anti-Akt phospho S473 Monoclonal antibody for IHC for mouse foreskin on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for mouse foreskin identification?
D. Collins
Verified customer
Asked: 2020-02-07
Answer
You can see on the product datasheet, MP00024 anti-Akt phospho S473 Monoclonal antibody has been tested for IF, IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse foreskin in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-02-07
Question
I see that the anti-Akt phospho S473 Monoclonal antibody MP00024 works with IHC, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-11-06
Answer
You can find protocols for IHC on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-11-06
Question
Is a blocking peptide available for product anti-Akt phospho S473 Monoclonal antibody (MP00024)?
Verified Customer
Verified customer
Asked: 2019-10-31
Answer
We do provide the blocking peptide for product anti-Akt phospho S473 Monoclonal antibody (MP00024). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-10-31
Question
Do you have a BSA free version of anti-Akt phospho S473 Monoclonal antibody MP00024 available?
Verified Customer
Verified customer
Asked: 2019-08-16
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Akt phospho S473 Monoclonal antibody MP00024 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-08-16
Question
Is this MP00024 anti-Akt phospho S473 Monoclonal antibody reactive to the isotypes of AKT1?
Verified Customer
Verified customer
Asked: 2019-06-04
Answer
The immunogen of MP00024 anti-Akt phospho S473 Monoclonal antibody is anti-AKT pS473 (MOUSE) Monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein, followed by hybridoma development. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2019-06-04
Question
We bought anti-Akt phospho S473 Monoclonal antibody for WB on adrenal gland in the past. I am using rat, and We are going to use the antibody for IHC next. My lab would like examining adrenal gland as well as cervix carcinoma in our next experiment. Could you please give me some suggestion on which antibody would work the best for IHC?
Verified Customer
Verified customer
Asked: 2019-05-22
Answer
I viewed the website and datasheets of our anti-Akt phospho S473 Monoclonal antibody and it appears that MP00024 has been tested on rat in both WB and IHC. Thus MP00024 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-05-22
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for foreskin using anti-Akt phospho S473 Monoclonal antibody MP00024. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-02-06
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-02-06
Question
We are currently using anti-Akt phospho S473 Monoclonal antibody MP00024 for rat tissue, and we are content with the IF results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on monkey tissues as well?
Verified Customer
Verified customer
Asked: 2018-06-25
Answer
The anti-Akt phospho S473 Monoclonal antibody (MP00024) has not been tested for cross reactivity specifically with monkey tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-06-25
Question
I have a question about product MP00024, anti-Akt phospho S473 Monoclonal antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2018-02-16
Answer
It is not recommended storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free MP00024 anti-Akt phospho S473 Monoclonal antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-02-16
Question
Our lab were happy with the WB result of your anti-Akt phospho S473 Monoclonal antibody. However we have seen positive staining in liver cytoplasm. using this antibody. Is that expected? Could you tell me where is AKT1 supposed to be expressed?
J. Carter
Verified customer
Asked: 2018-01-31
Answer
Based on literature, liver does express AKT1. Generally AKT1 expresses in cytoplasm. Regarding which tissues have AKT1 expression, here are a few articles citing expression in various tissues:
Adrenal gland, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 17081983, 18669648
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Foreskin, Pubmed ID: 1718748
Liver, Pubmed ID: 24275569
Muscle, and Ovary, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2018-01-31
Question
We have seen staining in rat muscle ovary. Any tips? Is anti-Akt phospho S473 Monoclonal antibody supposed to stain muscle ovary positively?
Verified Customer
Verified customer
Asked: 2017-06-12
Answer
According to literature muscle ovary does express AKT1. According to Uniprot.org, AKT1 is expressed in left adrenal gland, adrenal gland, muscle ovary, foreskin, cervix carcinoma, cervix carcinoma erythroleukemia, liver, among other tissues. Regarding which tissues have AKT1 expression, here are a few articles citing expression in various tissues:
Adrenal gland, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 17081983, 18669648
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Foreskin, Pubmed ID: 1718748
Liver, Pubmed ID: 24275569
Muscle, and Ovary, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2017-06-12
Question
Please see the WB image, lot number and protocol we used for foreskin using anti-Akt phospho S473 Monoclonal antibody MP00024. Please let me know if you require anything else.
W. Gonzalez
Verified customer
Asked: 2016-04-05
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2016-04-05
Question
Would MP00024 anti-Akt phospho S473 Monoclonal antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
R. Li
Verified customer
Asked: 2014-11-13
Answer
It shows on the product datasheet, MP00024 anti-Akt phospho S473 Monoclonal antibody as been validated on IHC. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2014-11-13
Question
We want to test anti-Akt phospho S473 Monoclonal antibody MP00024 on mouse foreskin for research purposes, then I may be interested in using anti-Akt phospho S473 Monoclonal antibody MP00024 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
G. Moore
Verified customer
Asked: 2013-01-31
Answer
The products we sell, including anti-Akt phospho S473 Monoclonal antibody MP00024, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2013-01-31