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Anti-UFM1 antibody [EPR4264(2)] ab109305 is a rabbit monoclonal antibody that is used in UFM1 western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR4264(2) is the most widely used clone for UFM1 on the market and is cited in >40 publications
- Specificity confirmed with UFM1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation


Images

Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (AB109305), expandable thumbnail
  • Western blot - Anti-UFM1 antibody [EPR4264(2)] (AB109305), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305), expandable thumbnail

Publications

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

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFIPFlow CytWB
Human
Tested
Not recommended
Tested
Not recommended
Tested
Mouse
Tested
Not recommended
Expected
Not recommended
Tested
Rat
Tested
Not recommended
Expected
Not recommended
Expected

Tested
Tested

Species

Mouse

Dilution info

1/250 - 1/1000

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/250 - 1/1000

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/250 - 1/1000

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/10 - 1/40

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000 - 1/2000

Notes

-

Species

Human

Dilution info

1/1000 - 1/2000

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Ubiquitin-like modifier which can be covalently attached via an isopeptide bond to lysine residues of substrate proteins as a monomer or a lysine-linked polymer (PubMed:15071506, PubMed:20018847, PubMed:29868776, PubMed:27653677). The so-called ufmylation, requires the UFM1-activating E1 enzyme UBA5, the UFM1-conjugating E2 enzyme UFC1, and the UFM1-ligase E3 enzyme UFL1 (PubMed:15071506, PubMed:20018847, PubMed:29868776, PubMed:27653677). Ufmylation is involved in reticulophagy (also called ER-phagy) induced in response to endoplasmic reticulum stress (PubMed:32160526). Ufmylation of TRIP4 regulates nuclear receptors-mediated transcription (PubMed:25219498).

Alternative names

Ubiquitin-fold modifier 1, BM-002, C13orf20, UFM1

Recommended products

Anti-UFM1 antibody [EPR4264(2)] ab109305 is a rabbit monoclonal antibody that is used in UFM1 western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR4264(2) is the most widely used clone for UFM1 on the market and is cited in >40 publications
- Specificity confirmed with UFM1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR4264(2)

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Storage information

Stable for 12 months at -20°C

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

UFM1 also called ubiquitin-fold modifier 1 is a small protein modifier with a mass of approximately 9 kDa. It is part of the ubiquitin-like protein family and primarily expressed in a wide range of tissues. UFM1 is involved in a unique form of post-translational modification known as UFMylation where it conjugates to substrate proteins altering their function and stability. This process plays a role in regulating cell cycle and apoptosis among other cellular processes.

Biological function summary

UFM1 functions as a signaling molecule that influences various cellular pathways. It acts as part of a complex with other enzymes including UBA5 UFC1 and UFL1 which are important for its conjugation process. This modification system plays a role in endoplasmic reticulum (ER) stress response by affecting proteins involved in ER-associated degradation (ERAD). Its interaction with these proteins helps cells manage stress and maintain homeostasis.

Pathways

UFM1 participates in the ER stress response pathway and is also connected to the mTOR signaling pathway. Both pathways are essential for cell growth and survival. Within these pathways UFM1 interacts with proteins like mLST8 in the mTOR pathway integrating signals that coordinate cell metabolism growth and apoptosis. The precise regulation of these pathways highlights UFM1's role in maintaining cellular equilibrium.

Associated diseases and disorders

UFM1 has been linked to cancer and congenital disorders. Altered UFMylation due to mutations in related genes can contribute to tumor development by affecting protein degradation and cellular stress responses. In hereditary conditions dysfunctional UFM1 activity can lead to rare disorders affecting metabolism and neurological function. Proteins like mLST8 which interact with UFM1 in pathways can also play a role in these disease mechanisms suggesting potential targets for therapeutic intervention.

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.

9 product images

  • Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    ab109305 immunoprecipitating UFM1. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at a dilution of 1/1000.

    Lane 1: 293 (Human embryonic kidney epithelial cell) whole cell lysate 10ug
    Lane 2: 293 (Human embryonic kidney epithelial cell) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109305 in 293 (Human embryonic kidney epithelial cell) whole cell lysate

    All lanes: Immunoprecipitation - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    Predicted band size: 9 kDa

  • Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    Mouse heart cell lysate Blocking and diluting buffer: 5% NFDM/TBST

    The band at around 28kDa might be UFM1 conjugates [PMID: 23152784].

    All lanes: Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/2000 dilution

    Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: Mouse spleen cell lysate at 20 µg

    Lane 4: Mouse heart cell lysate at 20 µg

    Lane 5: Rat spleen 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 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 9 kDa

    Observed band size: 28 kDa, 9 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    ab109305 staining UFM1 in rat kidney 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 1/1000. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    ab109305 staining UFM1 in mouse mouse liver 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) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    ab109305 staining UFM1 in human thyroid carcinoma 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) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    ab109305 staining UFM1 in human kidney 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 1/1000. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    All lanes: Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/1000 dilution

    Lane 1: Fetal kidney lysate at 10 µg

    Lane 2: 293T cell lysate at 10 µg

    Lane 3: SH-SY5Y cell lysate at 10 µg

    Lane 4: HepG2 cell lysate at 10 µg

    Predicted band size: 9 kDa

    Observed band size: 9 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab109305 at 1/250 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305), expandable thumbnail

    Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305)

    Western blot: Anti-UFM1 antibody [EPR4264(2)] (ab109305) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab109305 was shown to bind specifically to UFM1. A band was observed at 9/28 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in UFM1 knockout cell line. To generate this image, wild-type and UFM1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-UFM1 antibody [EPR4264(2)] (ab109305) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: UFM1 knockout HEK-293T cell lysate at 20 µg

    Lane 3: U-2 OS cell lysate at 20 µg

    Lane 4: HepG2 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 9 kDa, 28 kDa

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Product protocols

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