Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [8]
- Immunocytochemistry [2]
- Immunohistochemistry [3]
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- Product number
- PA5-78176 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SP3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Positive Control: HeLa, Jurkat, Jurkat_NE, Raji, NCI-H929 Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.2 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references A novel Cbx1, PurB, and Sp3 complex mediates long-term silencing of tissue- and lineage-specific genes.
Baksh SS, Pratt RE, Gomez J, Dzau VJ, Hodgkinson CP
The Journal of biological chemistry 2022 Jun;298(6):102053
The Journal of biological chemistry 2022 Jun;298(6):102053
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SP3 in whole cell lysate using 30 µg of protein. Samples were separated with 7.5 % SDS-PAGE and incubated with SP3 polyclonal antibody (Product # PA5-78176) using a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SP3 in A) Jurkat whole cell extract, B) Jurkat nuclear extract using 30 µg of protein. Samples were separated with 7.5% SDS-PAGE and incubated with SP3 polyclonal antibody (Product # PA5-78176) using a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SP3 in wild-type and SP3 knockout HeLa cells using 30 µg of protein. Samples were separated with 7.5% SDS-PAGE and incubated with SP3 polyclonal antibody (Product # PA5-78176) using a dilution of 1:1000 followed by HRP-conjugated anti-rabbit IgG and ECL substrate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of SP3 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a SP3 Polyclonal Antibody (Product # PA5-78176) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- SP3 Polyclonal Antibody detects SP3 protein by western blot analysis. A. 30 µg Jurkat whole cell extract. B. 30 µg Jurkat nuclear extract.7.5 % SDS-PAGE. SP3 Polyclonal Antibody (Product # PA5-78176) dilution: 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of SP3 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a SP3 Polyclonal Antibody (Product # PA5-78176) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SP3 was achieved by transfecting HeLa with SP3 specific siRNAs (Silencer® select Product # S13324, S13325). Western Blot analysis (Fig. a) was performed using whole cell extracts from the SP3 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with SP3 Polyclonal Antibody (Product # PA5-78176, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP (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 SP3.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using SP3 Polyclonal Antibody (Product # PA5-78176) and a ~85 kDa band corresponding to SP3 was observed across cell lines and tissues tested along with an uncharacterised band (*) at ~80 kDa. Nuclear extracts (30 µg lysate) of HeLa (Lane 1), RAJI (Lane 2), U-2 OS (Lane 3) HCT-116 (Lane 4), SW480 (Lane 5), HT-29 (Lane 6), NIH/3T3 (Lane 7), PC-12 (Lane 8); and tissue extract Mouse Testis (Lane 9) and Rat testis (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:2000 dilution) 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
- Immunofluorescent analysis of SP3 in HeLa cells. Samples were treated with 4% paraformaldehyde at RT for 5 min and incubated with SP3 polyclonal antibody (Product # PA5-78176) using a dilution of 1:500, followed by Hoechst.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- SP3 Polyclonal Antibody detects SP3 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: SP3 stained by SP3 Polyclonal Antibody (Product # PA5-78176) diluted at 1:500. Red: phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar= 10 µm.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- SP3 Polyclonal Antibody detects SP3 protein at nucleus on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. SP3 Polyclonal Antibody (Product # PA5-78176) dilution: 1:1,000. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- SP3 Polyclonal Antibody detects SP3 protein at nucleus on human ovarian carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human ovarian carcinoma. SP3 Polyclonal Antibody (Product # PA5-78176) dilution: 1:1,000. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- SP3 Polyclonal Antibody detects SP3 protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human lung cancer. SP3 stained by SP3 Polyclonal Antibody (Product # PA5-78176) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.