- PA5-28628 - Invitrogen Antibodies SCRIB antibody

Submitted references The Dimeric Form of HPV16 E6 Is Crucial to Drive YAP/TAZ Upregulation through the Targeting of hScrib.

Messa L, Celegato M, Bertagnin C, Mercorelli B, Alvisi G, Banks L, Palù G, Loregian A
Cancers 2021 Aug 13;13(16)

In-Depth Study of Transmembrane Mucins in Association with Intestinal Barrier Dysfunction During the Course of T Cell Transfer and DSS-Induced Colitis.

Breugelmans T, Van Spaendonk H, De Man JG, De Schepper HU, Jauregui-Amezaga A, Macken E, Lindén SK, Pintelon I, Timmermans JP, De Winter BY, Smet A
Journal of Crohn's & colitis 2020 Jul 30;14(7):974-994

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SCRIB Polyclonal Antibody (Product # PA5-28628). Sample (30 µg of whole cell lysate). Lane A: NIH-3T3. Lane B: JC. 5% SDS PAGE. SCRIB Polyclonal Antibody (Product # PA5-28628) diluted at 1:1,000. 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: Western Blot using SCRIB Polyclonal Antibody (Product # PA5-28628). Sample (30 µg of whole cell lysate). Lane A: HepG2. 5% SDS PAGE. SCRIB Polyclonal Antibody (Product # PA5-28628) diluted at 1:1,000. 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: Knockdown of SCRIB was achieved by transfecting HeLa cells with SCRIB specific siRNAs (Silencer® select Product # s23971, s23972). Western blot analysis (Fig. a) was performed using membrane enriched cell extracts from the SCRIB knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SCRIB Polyclonal Antibody (Product # PA5-28628, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 SCRIB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of Caco2 (Lane 1), T-47D (Lane 2), HeLa (Lane 3) and PC-3 (Lane 4). The blot was probed with Anti-SCRIB Polyclonal Antibody (Product # PA5-28628, 1:2000 dilution) 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 174 kDa band corresponding to SCRIB was observed across the cell lines tested except Caco-2 which showed no expression of the SRIB protein.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SCRIB Polyclonal Antibody detects SCRIB protein at cytoplasm on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded colon carcinoma. SCRIB Polyclonal Antibody (Product # PA5-28628) dilution: 1:250. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 The dimeric form of E6 localizes in the cytosol and takes part in the E6-hScrib complex formation. ( A ) Representative confocal image of the Myc-FLAG Proximity Ligation Assay (PLA) detecting E6 homodimerization (red fluorescence) in H1299 cells co-expressing Myc-E6 and FLAG-E6. The image represents the maximum intensity projection of a Z-stack acquired with a 2000x magnification. Scale bar: 5 mum. Left panels show the three-dimensional reconstruction of the indicated cell and both images show the same cell, with a 45 degrees rotation around the horizontal axis between the two images. Nuclei were stained with DRAQ5. Experimental controls are shown in Figure S3A,B . ( B ) Quantitative nuclear/cytoplasmic fluorescence (Fn/c) analysis of H1299 living cells overexpressing YFP-E6 or YFP-E6 Y43E/F47R. Data are presented as scatter-dot-plots of n YFP-E6 = 247 cells and n YFP-E6 Y43E/F47R = 251 cells. * p < 0.05 determined with the Mann-Whitney U test. Right panels show the representative images used for quantification. Scale bars: 20 mum. ( C ) CoIP/Western blot analysis (right) of H1299 cells overexpressing YFP-tagged and HA-tagged E6 proteins showing the binding of tagged E6 to hScrib through immunoprecipitation of endogenous hScrib. Control IP represents the extent of non-specific binding of each protein in the experiment. Left diagram provides a schematic representation of the result obtained with the experiment, with red and black circles on E6 models indicating amino

PA5-28628

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