- PA5-29275 - Invitrogen Antibodies PPM1A antibody

Submitted references LncRNA GAS5 attenuates fibroblast activation through inhibiting Smad3 signaling.

Tang R, Wang YC, Mei X, Shi N, Sun C, Ran R, Zhang G, Li W, Staveley-O'Carroll KF, Li G, Chen SY
American journal of physiology. Cell physiology 2020 Jul 1;319(1):C105-C115

Mesenchyme homeobox 1 mediates transforming growth factor-β (TGF-β)-induced smooth muscle cell differentiation from mouse mesenchymal progenitors.

Dong K, Guo X, Chen W, Hsu AC, Shao Q, Chen JF, Chen SY
The Journal of biological chemistry 2018 Jun 1;293(22):8712-8719

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PPM1A Polyclonal Antibody (Product # PA5-29275). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: A431. 10% SDS PAGE. PPM1A Polyclonal Antibody (Product # PA5-29275) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PPM1A was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a PPM1A Polyclonal Antibody (Product # PA5-29275) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PPM1A Polyclonal Antibody (Product # PA5-29275). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: A431. 10% SDS PAGE. PPM1A Polyclonal Antibody (Product # PA5-29275) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PPM1A Polyclonal Antibody (Product # PA5-29275). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 10% SDS PAGE. PPM1A Polyclonal Antibody (Product # PA5-29275) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PPM1A was achieved by transfecting HeLa cells with PPM1A specific siRNAs (Silencer® select Product # s10919, s10920). Western blot analysis (Fig. a) was performed using whole cell extracts from the PPM1A knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PPM1A Polyclonal Antibody (Product # PA5-29275, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (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 PPM1A. A 51KDa band corresponding to its isoform was observed along with an uncharacterized band at ~29KDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), A549 (Lane 3), PC-3 (Lane 4), A-431 (Lane 5), K-562 (Lane 6), Raji (Lane 7), Mouse kidney (Lane 8), and Mouse brain (Lane 9). The blot was probed with Anti-PPM1A Polyclonal Antibody (Product # PA5-29275, 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 42 kDa band corresponding to PPM1A was observed in all cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of PPM1A showing staining in the cytoplasm of A431 cells. A431 cells were fixed in ice-cold MeOH for 5 min and stained using a PPM1A polyclonal antibody (Product # PA5-29275) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PPM1A Polyclonal Antibody detects PPM1A protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in ice-cold MeOH for 5 min. Green: PPM1A protein stained by PPM1A Polyclonal Antibody (Product # PA5-29275) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PPM1A Polyclonal Antibody detects PPM1A protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in ice-cold MeOH for 5 min. Green: PPM1A protein stained by PPM1A Polyclonal Antibody (Product # PA5-29275) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PPM1A 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 1% BSA for 1 hour at room temperature. The cells were labeled with PPM1A Polyclonal Antibody (Product # PA5-29275) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization along with cytoplasm. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using PPM1A (Product # PA5-29275) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin immunoprecipitation analysis of PPM1A 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 PPM1A polyclonal antibody (Product # PA5-29275). 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

PA5-29275

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