Recombinant Anti-TRIM56 antibody [EPR10583] KO Tested (ab154862)

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Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR10583] to TRIM56
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
Knockout validated
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

Product name

Anti-TRIM56 antibody [EPR10583]
See all TRIM56 primary antibodies
Description

Rabbit monoclonal [EPR10583] to TRIM56
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide within Human TRIM56. The exact sequence is proprietary.
Database link: Q9BRZ2
Positive control
- IHC-P: Human brain, pancreas and colon cancer tissues. WB: HAP1, A549, A375, MCF7 and HeLa cell lysates. ICC/IF: MCF7 and HeLa cells Flow Cyt (intra): MCF7 cells
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
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR10583
Isotype

IgG
Research areas

Alternative Versions
- Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
- Alexa Fluor® 568 Anti-TRIM56 antibody [EPR10583] (ab312642)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human TRIM56 knockout A549 cell line (ab267062)
- Human TRIM56 knockout A549 cell line (ab267063)
KO cell lysates
- Human TRIM56 knockout A549 cell lysate (ab258249)
- Human TRIM56 knockout A549 cell lysate (ab258250)
Positive Controls
- HeLa whole cell lysate (ab29545)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab154862 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500 dilution.
WB		1/10000 - 1/50000. Predicted molecular weight: 81 kDa.
IHC-P		1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurifed use at 1/50 -1/100 dilution.
ICC/IF		1/70. For unpurified use at 1/100 - 1/250 dilution.

Notes
Flow Cyt (Intra) 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500 dilution.
WB 1/10000 - 1/50000. Predicted molecular weight: 81 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurifed use at 1/50 -1/100 dilution.
ICC/IF 1/70. For unpurified use at 1/100 - 1/250 dilution.

Function

E3 ubiquitin-protein ligase that mediates 'Lys-63'-linked polyubiquitination of TMEM173/STING, thereby playing a key role in innate immunity. TMEM173/STING 'Lys-63'-linked ubiquitination activates the production of type I interferon IFN-beta following detection of pathogen- and host-derived double-stranded DNA.
Sequence similarities

Belongs to the TRIM/RBCC family.
Contains 2 B box-type zinc fingers.
Contains 1 RING-type zinc finger.
Cellular localization

Cytoplasm.
Target information above from: UniProt accession Q9BRZ2 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 81844 Human
- SwissProt: Q9BRZ2 Human
- Unigene: 521092 Human
Alternative names
- A130009K11Rik antibody
- DKFZp667O116 antibody
- E3 ubiquitin-protein ligase TRIM56 antibody
- FLJ35608 antibody
- Gm452 antibody
- MGC37358 antibody
- OTTMUSP00000027392 antibody
- RING finger protein 109 antibody
- RNF109 antibody
- TRI56_HUMAN antibody
- Trim56 antibody
- Tripartite motif containing protein 56 antibody
- Tripartite motif-containing protein 56 antibody
see all

Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : A-375 (Human malignant melanoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 81 kDa
Observed band size: 88 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM56 antibody [EPR10583] (ab154862)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry (Intracellular) - Anti-TRIM56 antibody [EPR10583] (ab154862)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] (ab154862)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer tissue sections labeling TRIM56 with purified ab154862 at 1/1000 dilution (0.691 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : A-375 (Human malignant melanoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 81 kDa
Observed band size: 88 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/50000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : A375 (Human malignant melanoma epithelial cell) whole cell lysates

Lysates/proteins at 15 µg per lane.

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

Predicted band size: 81 kDa
Observed band size: 81 kDa
Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: TRIM56 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: A375 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab154862 (unpurifed) was shown to recognize TRIM56 when TRIM56 knockout samples were used, along with additional cross-reactive bands. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-TRIM56 antibody [EPR10583] (ab154862)

All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/10000 dilution ((unpurified))

Lane 1 : HeLa cell lysate
Lane 2 : A375 cell lysate
Lane 3 : MCF7 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 81 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] (ab154862)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling TRIM56 with ab154862 (unpurified) at 1/50.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] (ab154862)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling TRIM56 with ab154862 (unpurified) at 1/50.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM56 antibody [EPR10583] (ab154862)

Immunofluorescent analysis of MCF7 cells labeling TRIM56 with ab154862 (unpurified) at 1/100.
Anti-TRIM56 antibody [EPR10583] (ab154862)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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Publishing research using ab154862? Please let us know so that we can cite the reference in this datasheet.

ab154862 has been referenced in 6 publications.

Yang X et al. TRIM56 promotes malignant progression of glioblastoma by stabilizing cIAP1 protein. J Exp Clin Cancer Res 41:336 (2022). PubMed: 36471347
Yang P et al. G3BP1 Is a Tunable Switch that Triggers Phase Separation to Assemble Stress Granules. Cell 181:325-345.e28 (2020). PubMed: 32302571
Garcia-Moreno M et al. System-wide Profiling of RNA-Binding Proteins Uncovers Key Regulators of Virus Infection. Mol Cell 74:196-211.e11 (2019). PubMed: 30799147
Xue M et al. Regulation of estrogen signaling and breast cancer proliferation by an ubiquitin ligase TRIM56. Oncogenesis 8:30 (2019). PubMed: 31000690
Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
Fiskin E et al. Structural basis for the recognition and degradation of host TRIM proteins by Salmonella effector SopA. Nat Commun 8:14004 (2017). WB . PubMed: 28084320

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (6)

Customer reviews and Q&As