Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [4]
- Immunohistochemistry [10]
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- Product number
- PA5-95583 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SP1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- 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.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of SP1 in Lane 1: human K562 whole cell lysate, Lane 2: human A549 whole cell lysate, Lane 3: human HeLa whole cell lysate, Lane 4: human A431 whole cell lysate, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of SP1 was achieved by transfecting MCF7 with SP1 specific siRNAs (Silencer® select Product # S13320, S13319). Western blot analysis (Fig. a) was performed using Fresh lysates Whole cell extracts from the SP1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SP1 Polyclonal Antibody (Product # PA5-95583, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 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 SP1.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-SP1 Polyclonal Antibody (Product # PA5-95583) and a 95 kDa band along with uncharacterized band (*) corresponding to SP1 was observed across cell lines and tissue tested. Whole cell extracts (50 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3), HT-29 (Lane 4) and Rat Brain (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 :1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemistry analysis of SP1 in HeLa cell. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and developed with Strepavidin-Biotin-Complex/DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SP1 in A549 cell. Antigen retrieval was performed on the tissue using IHC enzyme antigen retrieval reagent and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by 488 conjugated goat anti-rabbit IgG using a 1:100 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescence analysis of SP1 in A431 cell. Antigen retrieval was performed on the tissue using IHC enzyme antigen retrieval reagent and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/1x10^6 cells dilution, followed by 488 conjugated goat anti-rabbit IgG using a 1:100 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SP1 was performed using 70 confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SP1 Polyclonal Antibody (Product # PA5-95583) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Alexa Fluor™ Plus 647 Phalloidin (Product # A30107, 1:2000 dilution). Panel d represents the merged image showing nuclear as well as cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al./Methods 115 (2017) 28–41).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded rat spleen tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded rat liver tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded rat small intestine tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded human intestinal cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded human lung cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded human esophagus squama cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded human ovary cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded human tonsil tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded mouse liver tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SP1 in paraffin-embedded rat brain tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with SP1 polyclonal antibody (Product # PA5-95583) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.