- PA5-78176 - Invitrogen Antibodies SP3 antibody

Baksh SS, Pratt RE, Gomez J, Dzau VJ, Hodgkinson CP
The Journal of biological chemistry 2022 Jun;298(6):102053

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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)
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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.

Supportive validation

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)
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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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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.

Supportive validation

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.

PA5-78176