Antibodies, Proteins, Kits and Reagents for Life Science

339 157 ₸ Product size

100 µl 339 157 ₸ 10 µl 139 232 ₸

Order now and get it on Tuesday March 02, 2021

Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR7315] to TRIM25/EFP
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-TRIM25/EFP antibody [EPR7315]
See all TRIM25/EFP primary antibodies
Description

Rabbit monoclonal [EPR7315] to TRIM25/EFP
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human TRIM25/EFP aa 100-200. The exact sequence is proprietary.
Database link: Q14258
Positive control
- Recombinant Human TRIM25/EFP protein (ab152814) can be used as a positive control in WB. HeLa and K562 cell lysates; Human breast tissue. Human pancreas tissue. Rat brain lysate. SP2/0 (mouse spleen) whole cell lysate.
General notes
This product was previously labelled as TRIM25

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
For more information see here.
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.

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.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR7315
Isotype

IgG
Research areas

Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TRIM25/EFP knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 71 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab167154 observed at 71 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab167154 was shown to specifically react with TRIM25/EFP in wild-type HAP1 cells as signal was lost in TRIM25/EFP knockout cells. Wild-type and TRIM25/EFP knockout samples were subjected to SDS-PAGE. Ab167154 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/50000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 71 kDa

Blocking and diluting buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

ab167154 staining TRIM25/EFP in human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

ab167154 staining TRIM25/EFP in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a tubulin counterstain at a dilution of 1/200 and DAPI was used as a nuclear counterstain.
Flow Cytometry - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling TRIM25/EFP with purified ab167154 at 1/90 dilution(10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/2000 dilution + SP2/0 (mouse spleen) whole cell lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 71 kDa
Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.

Blocking and diluting buffer: 5% NFDM /TBST
Immunoprecipitation - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

ab167154 immunoprecipitating TRIM25/EFP. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab167154 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate.
Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution + Rat brain lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 71 kDa
Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.

Blocking and diluting buffer: 5% NFDM/TBST
Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : K562 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 71 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling TRIM25/EFP with ab167154 at 1/100 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Immunofluorescent analysis of HeLa cells labeling TRIM25/EFP with ab167154 at 1/100 dilution.
Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

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