PA5-27145

antibody from Invitrogen Antibodies

Targeting: FOXO3 AF6q21, FKHRL1, FOXO2, FOXO3A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-27145

Provider

Invitrogen Antibodies

Product name

FOXO3A Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: 293T, Mouse brain, C2C12, Jurkat, Raji. Predicted reactivity: Mouse (100%), Rat (100%), Pig (100%), Sheep (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.56 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Lipid Droplet-Derived Monounsaturated Fatty Acids Traffic via PLIN5 to Allosterically Activate SIRT1.

Najt CP, Khan SA, Heden TD, Witthuhn BA, Perez M, Heier JL, Mead LE, Franklin MP, Karanja KK, Graham MJ, Mashek MT, Bernlohr DA, Parker L, Chow LS, Mashek DG
Molecular cell 2020 Feb 20;77(4):810-824.e8

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Impact of first-line cancer treatment on the follicle quality in cryopreserved ovarian samples from girls and young women.

Pampanini V, Wagner M, Asadi-Azarbaijani B, Oskam IC, Sheikhi M, Sjödin MOD, Lindberg J, Hovatta O, Sahlin L, Björvang RD, Otala M, Damdimopoulou P, Jahnukainen K
Human reproduction (Oxford, England) 2019 Sep 29;34(9):1674-1685

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of FOXO3A using 30 µg of 293T lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a FOXO3A polyclonal antibody (Product # PA5-27145) at a dilution of 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of FOXO3 was achieved by transfecting MCF7 cells with FOXO3 specific validated siRNAs (Silencer® select Product # S5261, S5260). Western blot analysis (Fig. a) was performed using whole cell extracts (1% SDS) from the FOXO3 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with FOXO3 Monoclonal Antibody (Product #, PA5-27145 #, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Decrease in signal upon siRNA mediated knockdown confirms that antibody is specific to FOXO3.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF7 (Lane 1) and Hep G2 (Lane 2). The blot was probed with Anti-FOXO3 Rabbit Polyclonal Antibody (Product # PA5-27145, 1 µg/mL) 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 71 kDa band corresponding to FOXO3A was detected across the cell lines tested.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using FOXO3A Polyclonal Antibody (Product # PA5-27145). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 7.5% SDS PAGE. FOXO3A Polyclonal Antibody (Product # PA5-27145) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using FOXO3A Polyclonal Antibody (Product # PA5-27145). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with FOXO3A Polyclonal Antibody (Product # PA5-27145) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of FOXO3A was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: FOXO3A Polyclonal Antibody (Product # PA5-27145) diluted at 1:1000. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 Exposure to chemotherapy before ovarian tissue cryopreservation does not increase activation of ovarian follicles . ( A ) Box plots showing a significantly smaller proportion of growing follicles in ovarian tissue of treated ( n = 14) and untreated ( n = 13) patients. For each patient, the number of intact growing (intermediary and primary) follicles was divided by the total number of intact and atretic follicles (primordial, intermediary and primary) *** ( P < 0.001). ( B ) Immunofluorescence of ovarian tissue from a subset of treated ( n = 3) and untreated ( n = 5) patients labeled with fork head box O3A (FOXO3A) antibody shows no differences between the two groups. FOXO3A is located in the nuclei of oocytes and granulosa cells of both treated and untreated patients indicating non-activated follicles. Representative images of ovarian tissue sections are shown.