- PA5-80339 - Invitrogen Antibodies ANP32A antibody

Gebhart NN, Hardy RW, Sokoloski KJ
PloS one 2020;15(8):e0238254

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PHAP I in Lane A: HEK293 Whole Cell Lysate (30 µg), Lane B: Raji Whole Cell Lysate (30 µg), Lane C: HeLa Whole Cell Lysate (30 µg), Lane D: Jurkat Whole Cell Lysate (30 µg). Samples were probed using a PHAP I Polyclonal Antibody (Product # PA5-80339) at a 1:500 dilution, followed by a Goat Anti-Rabbit IgG (H+L), Dylight 800 Secondary Antibody at a 1:10000 dilution. Western blot was performed under reducing conditions. Predicted band size:28 kDa. Observed band size:28 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PHAP I in Lane A: ANP32A konckout Hela Whole Cell Lysate (20 µg), Lane B: Hela Whole Cell Lysate (20 µg). Samples were probed using a PHAP I Polyclonal Antibody (Product # PA5-80339) at a 1:500 dilution, followed by a Goat Anti-Rabbit IgG (H+L), HRP Secondary Antibody at a 1:10000 dilution. Western blot was performed under reducing conditions. Predicted band size:28 kDa. Observed band size:28 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PHAP I Polyclonal Antibody (Product # PA5-80339) at 1:500 dilution. Lane A: PHAP I konckout Hela Whole Cell Lysate, Lane B: Hela Whole Cell Lysate. Lysates/proteins at 20 μg per lane. Secondary antibody: Goat Anti-Rabbit IgG (H+L)/HRP at 1:10,000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size: 28 kDa. Observed band size: 28 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PHAP I Polyclonal Antibody (Product # PA5-80339) at 1:500 dilution. Lane A: HEK293 Whole Cell Lysate, Lane B: Raji Whole Cell Lysate, Lane C: HeLa Whole Cell Lysate, Lane D: Jurkat Whole Cell Lysate. Lysates/proteins at 30 μg per lane. Secondary Goat Anti- RabbitIgG H&L (DyLight™ 800) at 1:10,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 28 kDa. Observed band size: 28 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of PHAP I in HeLa cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with PHAP I Polyclonal Antibody (Product # PA5-80339, 1:1,000) at 4°C overnight. Then cells were stained with the Alexa Fluor®488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue). Positive staining was localized to nucleus.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of PHAP I in HeLa cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with PHAP I Polyclonal Antibody (Product # PA5-80339, 1:1,000) at 4°C overnight. Then cells were stained with the Alexa Fluor®488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue). Positive staining was localized to nucleus.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of human PHAP I in human kidney with PHAP I Polyclonal Antibody (Product # PA5-80339, 1:1,000, formalin-fixed paraffin embedded sections).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of human PHAP I in human skeletal muscle with PHAP I Polyclonal Antibody (Product # PA5-80339, 1:1,000, formalin-fixed paraffin embedded sections).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PHAP I Immunoprecipitation using: Lane A: 0.5 mg Jurkat Whole Cell Lysate, Lane B: 0.5 mg Hela Whole Cell Lysate, Lane C: 0.5 mg HepG2 Whole Cell Lysate 4 µL with PHAP I Polyclonal Antibody (Product # PA5-80339) and 15 µL of 50 % Protein G agarose. Primary antibody: PHAP I Polyclonal Antibody, at 1:100 dilution. Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 29 kDa. Observed band size: 29 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 9 The validation of select CLAMP identified common interactants. 293HEK cells were infected with SINV, CHIKV, or VEEV at an MOI of 10 PFU/Cell and processed as described in the materials and methods. Briefly, the infected cells were cross-linked prior to the generation of whole cell lysates. After clarification via centrifugation the cross-linked lysates were immunoprecipitated with either anti-hnRNP K (Panel A), anti-hnRNP A1 (Panel B), or anti-ANP32a antibodies (Panel C). Nonspecific control immunoprecipitations utilizing an anti-IL1 antibody were conducted in parallel. After immunoprecipitation the cross-links were reversed, and the co-immunoprecipitated RNAs were used as input materials for the synthesis of random hexamer primed cDNAs. The positive sense viral RNA species of SINV, CHIKV, and VEEV were then assayed using qRT-PCR and plotted as the fold abundance of the viral RNA relative to the control immunoprecipitations. The quantitative data shown is the means of three independent biological replicates, and the error bar represents the standard deviation of the means.

PA5-80339