Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [6]
- Flow cytometry [1]
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- Product number
- PA5-95056 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- 14-3-3 sigma Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Persistence of a regeneration-associated, transitional alveolar epithelial cell state in pulmonary fibrosis.
Kobayashi Y, Tata A, Konkimalla A, Katsura H, Lee RF, Ou J, Banovich NE, Kropski JA, Tata PR
Nature cell biology 2020 Aug;22(8):934-946
Nature cell biology 2020 Aug;22(8):934-946
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of 14-3-3 sigma in Lane 1: HeLa whole cell lysate (40 µg), Lane 2: A549 whole cell lysate (40 µg), Lane 3: A549 whole cell lysate (40 µg), Lane 4: COLO320 whole cell lysate (40 µg), Lane 5: SE620 whole cell lysate (40 µg). Samples were incubated with 14-3-3 sigma polyclonal antibody (Product # PA5-95056).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SFN in, Lane 1: human Hela whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: mouse brain tissue lysates. 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 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with 14-3-3 sigma Polyclonal Antibody (Product # PA5-95056) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit 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. A specific band was detected for SFN at approximately 28KD. The expected band size for SFN is at 28KD.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of 14-3-3 protein sigma was achieved by transfecting A549 with 14-3-3 protein sigma specific siRNAs (Silencer® select Product # s5960, s5961). Western blot analysis (Fig. a) was performed using Whole cell extracts from the 14-3-3 protein sigma knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with 14-3-3 sigma Polyclonal Antibody (Product # PA5-95056, 1:4000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 protein sigma.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-14-3-3 sigma Polyclonal Antibody (Product # PA5-95056) and a 30 kDa band corresponding to 14-3-3 protein sigma, along with an uncharacterized band (*), was observed across cell lines and tissues tested. Whole cell extracts (40 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), A549 (Lane 3), MCF7 (Lane 4) and tissue extract of Mouse Stomach (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.20 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of SFN using anti-SFN antibody (Product # PA5-95056) . SFN was detected in a section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-SFN antibody (Product # PA5-95056) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of SFN using anti-SFN antibody (Product # PA5-95056). SFN was detected in a section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-SFN antibody (Product # PA5-95056) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of 14-3-3 sigma in human esophagus squama cancer tissue. Samples were incubated with 14-3-3 sigma polyclonal antibody (Product # PA5-95056).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of 14-3-3 sigma in human tonsil tissue. Samples were incubated with 14-3-3 sigma polyclonal antibody (Product # PA5-95056).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of 14-3-3 sigma in human tonsil tissue. Samples were incubated with 14-3-3 sigma polyclonal antibody (Product # PA5-95056).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SFN in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-SFN antibody (Product # PA5-95056) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SFN in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-SFN antibody (Product # PA5-95056) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SFN in paraffin-embedded section of human oesophagus squama cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-SFN antibody (Product # PA5-95056) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of 14-3-3 sigma in U2OS cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with 14-3-3 sigma Polyclonal Antibody (Product # PA5-95056) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.