Anti-PRMT5 antibody [EPR5772] (ab109451)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5772] to PRMT5
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-PRMT5 antibody [EPR5772]
See all PRMT5 primary antibodies -
Description
Rabbit monoclonal [EPR5772] to PRMT5 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P MouseHumanIP MouseWB MouseHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, HepG2, HeLa, and NIH/3T3 cell lysates; mouse and rat brain tissue lysate. ICC/IF: HepG2 and HeLa cells. IHC-P: Human infiltrating duct carcinoma of breast tissue, mouse liver tissue. Flow Cyt: HeLa cells. IP: Mouse brain cell.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 6.70 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5772 -
Isotype
IgG -
Research areas
Images
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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
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All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/50000 dilution (purified)
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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Expression and localization of candidate epigenetic reprogramming factors in human embryos.
Human blastocysts were incubated with primary antibodies for (Panel B) PRMT5 (green), the nuclear DNA stained with DAPI (blue) and visualized by multi-channel confocal microscopy. Note that the expression of PRMT5 primarily localized to the ICM of human blastocysts (indicated by white arrows).
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Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified)
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling 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: Secondary antibody Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/400).
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ICC/IF image of ab109451 (unpurified) stained HepG2 (Human liver hepatocellular carcinoma cell line) cells.
The cells were fixed with 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize 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 1.43 µM.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling 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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue sections labeling 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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa (Human liver hepatocellular carcinoma cell line) 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1 µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
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Equilibrium disassociation constant (KD)
Click here to learn more about KD -