- PA5-28218 - Invitrogen Antibodies PPP1CA antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PPP1A using 30 µg of A) 293T and B) A431 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a PPP1A polyclonal antibody (Product # PA5-28218) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of PP1 alpha was performed by separating 30 µg of whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a PP1 alpha Polyclonal Antibody (Product # PA5-28218) 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)
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Experimental details: Western Blot using PP1 alpha Polyclonal Antibody (Product # PA5-28218). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with PP1 alpha Polyclonal Antibody (Product # PA5-28218) 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)
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Experimental details: Western Blot to detect PP1 alpha from U2OS cells treated with control siRNA or PP1 alpha siRNA, using PP1 alpha Polyclonal Antibody (Product # PA5-28218) at 1:1,000 dilution. Nuclear matrix protein p84 is a loading control, blotted with p84 antibody (clone 5E10) (Product # MA1-23261) at 1:5,000 dilution. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PP1 alpha Polyclonal Antibody detects PPP1A protein by western blot analysis. Rat tissue extracts (50 µg) was separated by 10% SDS-PAGE, and the membrane was blotted with PP1 alpha Polyclonal Antibody (Product # PA5-28218) diluted by 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of PPP1A in paraformaldehyde-fixed HeLa cells using a PPP1A polyclonal antibody (Product # PA5-28218) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded SAS xenograft, using PPP1A (Product # PA5-28218) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Chromatin immunoprecipitation analysis of PPP1A was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a PPP1A polyclonal antibody (Product # PA5-28218). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify exon 3-3 of human Bcl-x (hBcl-x) or exon 1 of human NR4A3 (hNR4A3). PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. Schematic representations of the Bcl-x and NR4A3 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions). Regions amplified by Bcl-x and NR4A3 primers are represented by black bars. Data courtesy of the Innovators Program.

PA5-28218