- PA5-78323 - Invitrogen Antibodies PRMT5 antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PRMT5 in Raji cells using 30 µg of protein. Samples were separated with 7.5% SDS-PAGE and incubated with PRMT5 polyclonal antibody (Product # PA5-78323) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PRMT5 Polyclonal Antibody (Product # PA5-78323). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with PRMT5 Polyclonal Antibody (Product # PA5-78323) 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 PRMT5 was achieved by transfecting HeLa with PRMT5 specific siRNAs (Silencer® select Product # s20376). Western blot analysis (Fig. a) was performed using whole cell extracts from the PRMT5 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PRMT5 Monoclonal Antibody (Product # PA5-78323, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant 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 PRMT5.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using PRMT5 Polyclonal Antibody (Product # PA5-78323) and bands of 66, 67 kDa corresponding to PRMT5 was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF-7 (Lane 2), T47D (Lane 3), Hep G2 (Lane 4), PC-3 (Lane 5) and NIH/3T3 (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) 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)
Main image
Experimental details: Immunofluorescence analysis of PRMT5 was performed using 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 2% BSA for 1 hour at room temperature. The cells were labeled with PRMT5 Polyclonal Antibody (Product # PA5-78323) at 1:200 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034, 1:2000 dilution) for 45 minutes at room temperature (Panel a: green) in HeLa cells. 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 of HeLa cells showing nucleoplasm and cytosol for PRMT5. 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 MCF7 xenograft, using PRMT5 (Product # PA5-78323) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PRMT5 Polyclonal Antibody detects PRMT5 protein at cytoplasm and nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. PRMT5 stained by PRMT5 Polyclonal Antibody (Product # PA5-78323) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin immunoprecipitation analysis of PRMT5 in K562 chromatin extract using 5 µg of sample. Immunoprecipitation was performed using PRMT5 polyclonal antibody (Product # PA5-78323). Analysis of the precipitated DNA was done by PCR using primer targeting γ-globin promoter.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Cross-linked ChIP was performed with K562 chromatin extract and 5 µg of either control rabbit IgG or PRMT5 Polyclonal Antibody (Product # PA5-78323). The precipitated DNA was detected by PCR with primer set targeting to gamma-globin promoter.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PRMT5 Polyclonal Antibody immunoprecipitates PRMT5 protein in IP experiments. IP samples: Raji whole cell extract. A. 35 µg Raji whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of PRMT5 protein by 4 µg PRMT5 Polyclonal Antibody (Product # PA5-78323). 7.5 % SDS-PAGE. The immunoprecipitated PRMT5 protein was detected by PRMT5 Polyclonal Antibody (Product # PA5-78323) diluted at 1:500.

PA5-78323