Antibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Flow cytometry [1]
- Other assay [4]
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- Product number
- MA5-14916 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AKT Pan Monoclonal Antibody (J.314.4)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Antibody clone number
- J.314.4
- Concentration
- 35 µg/mL
Submitted references CDKL3 promotes osteosarcoma progression by activating Akt/PKB.
Extracellular matrix promotes proliferation, migration and adhesion of airway smooth muscle cells in a rat model of chronic obstructive pulmonary disease via upregulation of the PI3K/AKT signaling pathway.
He A, Ma L, Huang Y, Zhang H, Duan W, Li Z, Fei T, Yuan J, Wu H, Liu L, Bai Y, Dai W, Wang Y, Li H, Sun Y, Wang Y, Wang C, Yuan T, Yang Q, Tian S, Dong M, Sheng R, Xiang D
Life science alliance 2020 May;3(5)
Life science alliance 2020 May;3(5)
Extracellular matrix promotes proliferation, migration and adhesion of airway smooth muscle cells in a rat model of chronic obstructive pulmonary disease via upregulation of the PI3K/AKT signaling pathway.
Wang Z, Li R, Zhong R
Molecular medicine reports 2018 Sep;18(3):3143-3152
Molecular medicine reports 2018 Sep;18(3):3143-3152
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Supportive validation
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- Knockdown of AKT Pan was achieved by transfecting A-431 cells with AKT Pan specific siRNAs (Silencer® select Cat # s1215, s19429, s659).Western blot analysis (Fig a) was performed using whole cell lysates from the AKT Pan knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with anti-AKT Pan Monoclonal Antibody (Product MA5-14916, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody HRP conjugate (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Reduction in signal upon siRNA mediated knock down confirms that antibody is specific to AKT Pan.
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- Western blot analysis was performed on whole cell extracts (30 µg lysate) of LNCap (Lane 1), PC-3 (Lane 2), U-87 MG (Lane 3), SH-SY5Y (Lane 4), T98G (Lane 5), A-431 (Lane 6), HCT 116 (Lane 7), H1975 (Lane 8), A549 (Lane 9), PC-12 (Lane 10) and NIH/3T3 (Lane 11). The blot was probed with Anti-AKT Pan Monoclonal Antibody (Product # MA5-14916, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 56 kDa band corresponding to AKT Pan was observed across the cell lines tested.
Supportive validation
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- Immunofluorescent analysis of Akt (pan) in C2C12 cells, PI3K-Inhibitor-treated, using an Akt (pan) monoclonal antibody (Product # MA5-14916) (green). Actin filaments are labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.
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- Immunofluorescent analysis of Akt (pan) in C2C12 cells, insulin-treated, using an Akt (pan) monoclonal antibody (Product # MA5-14916) (green). Actin filaments are labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.
Supportive validation
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- Immunohistochemical analysis of Akt (pan) in paraffin-embedded human melanoma using an Akt (pan) monoclonal antibody (Product # MA5-14916).
Supportive validation
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- Flow cytometric analysis of Akt (pan) in Jurkat cells using a Akt (pan) monoclonal antibody (Product # MA5-14916) (blue) compared to a nonspecific negative control antibody (red).
Supportive validation
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- Figure 7. Effects of extracellular matrix components of ASMCs in rat models of chronic obstructive pulmonary disease are mediated by the PI3K/AKT signaling pathway. The wound area was (A) visualized and (B) calculated to evaluate motility of ASMCs following combined treatment with COL-3 and PI3K inhibitor LY294002 or activator IGF1. (C) Western blot analysis of components of the PI3K/AKT signaling pathway in ASMCs following combined treatment with COL-3 and PI3K inhibitor LY294002 or activator IGF1. Data are presented as the mean +- standard deviation in triplicate. **P
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- Figure 5. Cyclin-dependent kinase-like 3 (CDKL3) critically regulates Akt activation in osteosarcoma (OS). (A) Akt and mTOR activation under regular or starvation conditions in parental and CDKL3-KO U2OS cells. Upon CDKL3 KO, Akt and mTORC1 activation were significantly alleviated. The numbers below each blotting strip is intensity quantification by ImageJ. (B) The expression patterns of mTORC1 and FoxO downstream target genes confirm that CDKL3 regulates both pathways at the transcription level. Cells were cultured under the regular growth condition. (C) Overexpression of CDKL3 in U2OS cells causes hyper-activation of Akt in the presence and absence of FBS. (D) General working mechanism of rapamycin, AZD5363, and MK-2206. (E) U2OS cell growth under different conditions (n = 3). (F) Co-immunoprecipitation (Co-IP) reveals the physical interaction between HA-CDKL3 and Myc-Akt1. (G) Endogenous CDKL3 coexists with endogenous Akt shown by co-IP. (H) Schematic diagrams of Akt1 constructs for mapping. (I) Co-IP of HA-CDKL3 with different Akt1 constructs shows that the Akt kinase domain is indispensable for CDKL3 interaction. (I, J) Reverse IP of CDKL3 and Akt1 confirms the findings in (I). Error bars indicate SD (n = 3). * P < 0.05 parental DMSO versus CDKL3 -/- DMSO, or parental rapamycin, or parental AZD5363, or parental MK-2206; ** P < 0.01 CDKL3 -/- DMSO versus CDKL3 -/- rapamycin, or CDKL3 -/- AZD5363, or CDKL3 -/- MK-2206; ## P < 0.01 parental MK-2206 versus CDKL3 -/- MK-2206.
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- Figure 6. Cyclin-dependent kinase-like 3 (CDKL3) defines poor prognosis and correlates with Akt phosphorylation in clinic. (A, B) Kaplan-Meier plots of overall survival (A) and metastasis-free survival (B) of 152 osteosarcoma (OS) patients, stratified by CDKL3 levels (- represents negative staining; +, ++, and +++ represent weak, intermediate, and strong staining, respectively). (C) Representative immunohistochemistry (IHC) images of OS biopsies with different levels of CDKL3 expression on an OS microarray containing 152 primary OS tissues samples. Scale bar = 500 mum. (D) Representative HE and IHC images of OS biopsies stained by CDKL3 and Ki67. Scale bar = 100 mum. (E) Quantitative analysis of Ki67 expression in OS patients with different levels of CDKL3 expression. (F) HE and IHC staining images of CDKL3, AKT, pAKT (p308), pAKT (ser473), and LC3 in representative CDKL3-positive and CDKL3-negative patients. Scale bar = 100 mum. (G) Western blot detection of CDKL3 and Akt phosphorylation in OS tissues and adjacent non-tumor tissues from three different patients. (H) Putative underlying mechanism that CDKL3 promotes OS progression. Overexpression of CDKL3 leads to increased phosphorylation of Akt, followed by governing mTORC1 and FoxO activities, likely independent of functions of PDK1, growth factors, or relevant receptors; this may inhibit autophagy and eventually promote OS development. * P < 0.05, ** P < 0.01, *** P < 0.001, two-tailed t test. Source data are available fo
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- Figure S8. HA-CDKL3 co-immunoprecipitates endogenous Akt. Source data are available for this figure. Source Data for Figure S8