Anti-PRMT5 antibody [EPR5772] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PRMT5 (green) with purified ab109451 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control : primary antibody (1/50) and secondary antibody Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/400).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109451).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

ICC/IF image of ab109451 (unpurified) stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109451, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109451).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

Overlay histogram showing HeLa cells stained with ab109451 (unpurified, red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109451, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109451).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue sections labelling PRMT5 with purified ab109451 at 1/100. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109451).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labelling PRMT5 with purified ab109451 at 1/100. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109451).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

This data was developed using ab109451, the same antibody clone in a different buffer formulation.PRMT5 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab109451 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab109451 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : ab109451 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab109451 in mouse brain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 100 seconds

All lanes:

Immunoprecipitation - Anti-PRMT5 antibody [EPR5772] (<a href='/en-us/products/primary-antibodies/prmt5-antibody-epr5772-ab109451'>ab109451</a>)

Predicted band size: 36 kDa,73 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PRMT5 antibody [EPR5772] - BSA and Azide free (AB215364)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.