Anti-TRIM32 antibody [EPR29683-521] (ab322979)

Rabbit Monoclonal TRIM32 antibody. Suitable for ICC/IF, IHC-P, WB and reacts with Human, Mouse samples.

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Images

Western blot - Anti-TRIM32 antibody [EPR29683-521] (AB322979), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IHC-P	WB	IHC-Fr
Human	Tested	Expected	Tested	Not recommended
Mouse	Tested	Tested	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -
Species Mouse	Dilution info 1/50	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human	Dilution info -	Notes -

Target data

See full target information Trim32

Function

E3 ubiquitin ligase that plays a role in various biological processes including neural stem cell differentiation, innate immunity, inflammatory resonse and autophagy (PubMed:14578165, PubMed:37415157). Plays a role in virus-triggered induction of IFN-beta and TNF-alpha by mediating the ubiquitination of STING1. Mechanistically, targets STING1 for 'Lys-63'-linked ubiquitination which promotes the interaction of STING1 with TBK1. Regulates bacterial clearance and promotes autophagy in Mycobacterium tuberculosis-infected macrophages (By similarity). Negatively regulates TLR3/4-mediated innate immune and inflammatory response by triggering the autophagic degradation of TICAM1 in an E3 activity-independent manner (PubMed:28898289). Plays an essential role in oxidative stress induced cell death by inducing loss of transmembrane potential and enhancing mitochondrial reactive oxygen species (ROS) production during oxidative stress conditions. Ubiquitinates XIAP and targets it for proteasomal degradation. Ubiquitinates DTNBP1 (dysbindin) and promotes its degradation. May ubiquitinate BBS2 (By similarity). Ubiquitinates PIAS4/PIASY and promotes its degradation in keratinocytes treated with UVB and TNF-alpha (By similarity). Also acts as a regulator of autophagy by mediating formation of unanchored 'Lys-63'-linked polyubiquitin chains that activate ULK1: interaction with AMBRA1 is required for ULK1 activation. Positively regulates dendritic branching by promoting ubiquitination and subsequent degradation of the epigenetic factor CDYL (By similarity). Under metabolic stress and phosphorylation by CHK2, mediates 'Lys-63'-linked ubiquitination of ATG7 at 'Lys-41' to initiate autophagy (By similarity).

Additional Targets

TRIM32

Alternative names

E3 ubiquitin-protein ligase TRIM32, RING-type E3 ubiquitin transferase TRIM32, Tripartite motif-containing protein 32

Rabbit Monoclonal TRIM32 antibody. Suitable for ICC/IF, IHC-P, WB and reacts with Human, Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Clone number: EPR29683-521
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for human IHC-P.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: HL-60
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979) at 1/1000 dilution
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: U-118 MG (human brain glioblastoma cell) whole cell lysate at 20 µg
Lane 3: HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg
Lane 4: RD (human muscle mtinucleated spindle-shaped cell) whole cell lysate at 20 µg
Lane 5: HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 72 kDa, 36 kDa
Exposure time: 48s
Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: NIH/3T3 transfected with siRNA specifically targeting TRIM32 whole cell lysate at 20 µg
Lane 3: A20 (mouse reticum sarcoma B lymphocyte) whole cell lysate at 20 µg
Lane 4: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg
Lane 5: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 72 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: spleen, skeletal muscle, liver(PMID: 19349376)
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
Exposure time: Lane1-3: 10 seconds, lane4-5: 26seconds
All lanes: Western blot - Anti-TRIM32 antibody [EPR29683-521] (ab322979) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse spleen tissue lysate at 20 µg
Lane 3: Mouse skeletal muscle tissue lysate at 20 µg
Lane 4: Mouse testis tissue lysate at 20 µg
Lane 5: Mouse liver tissue lysate
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 72 kDa, 36 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling TRIM32 with ab322979 at 1/500 (0.982 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression tissue: No staining in mouse liver (PMID: 14578165).

The section was incubated with ab322979 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling TRIM32 with ab322979 at 1/500 (0.982 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining in mouse glioblastoma.

The section was incubated with ab322979 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunohistochemical analysis of paraffin-embedded Mouse squamous cell carcinoma tissue labeling TRIM32 with ab322979 at 1/500 (0.982 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining in mouse squamous cell carcinoma.

The section was incubated with ab322979 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling TRIM32 with ab322979 at 1/500 (0.982 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining in mouse testis.

The section was incubated with ab322979 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TRIM32 with ab322979 at 1/500 (0.982 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining in mouse cerebrum.

The section was incubated with ab322979 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-1080 (human fibrosarcoma epithelial cell) cells labelling TRIM32 with ab322979 at 1/50 (9.82 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in HT-1080 cells and weak cytoplasmic staining in HL-60 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse splenocyte cells labelling TRIM32 with ab322979 at 1/50 (9.82 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing no staining in mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression cells: Mouse spleen cell (PMID: 19349376). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-TRIM32 antibody [EPR29683-521] (ab322979)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling TRIM32 with ab322979 at 1/50 (10.14 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: ab322979 at a 1/50 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at a 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution.