Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-23507 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- 14-3-3 sigma Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Reactivity
- Human, Bovine, Canine
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of 14-3-3 sigma was achieved by transfecting A549 cells with 14-3-3 sigma specific validated siRNAs (Silencer® select Product # s5960, s5961). Western blot analysis (Fig. a) was performed using whole cell extracts from the 14-3-3 sigma knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with 14-3-3 sigma Polyclonal Antibody (Product # PA5-23507, 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 knock down confirms that antibody is specific to 14-3-3 sigma.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts of A-431 (Lane 1), A-375 (Lane 2), U-87 MG (Lane 3) and HCT 116 (Lane 4). The blot was probed with Anti-14-3-3 sigma antibody (Product # PA5-23507, 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 28 kDa band corresponding to 14-3-3 sigma was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of 14-3-3 sigma in human brain tissue in the 1) absence and 2) presence of immunizing peptide, 3) mouse brain and 4) rat brain tissue. Samples were incubated in 14-3-3 sigma polyclonal antibody (Product # PA5-23507) using a dilution of 3 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of 14-3-3 sigma in human A431 cell lysate in the 1) absence and 2) presence of immunizing peptide, and 3) HeLa cell lysate. Samples were incubated in 14-3-3 sigma polyclonal antibody (Product # PA5-23507) using a dilution of 3 µg/mL.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of 4EBP1 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with 14-3-3 sigma Rabbit Polyclonal Antibody (Product # PA5-23507) at 5 µg/mL concentration in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of 14-3-3 sigma in formalin-fixed, paraffin-embedded human tonsil (squamous epithelium). Samples were incubated in 14-3-3 sigma polyclonal antibody (Product # PA5-23507) using a dilution of 10 µg/mL.