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Rabbit Recombinant Monoclonal SHARPIN antibody. Suitable for WB and reacts with Human, Mouse samples.

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Images

Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891), expandable thumbnail
  • Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891), expandable thumbnail
  • Western blot - Anti-SHARPIN antibody [EPR29120-74] (AB321891), 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

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-FrICC/IFIHC-P
Human
Tested
Not recommended
Not recommended
Not recommended
Mouse
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse

Dilution info

-

Notes

-

Target data

Function

Component of the LUBAC complex which conjugates linear polyubiquitin chains in a head-to-tail manner to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation (PubMed:21455173, PubMed:21455180, PubMed:21455181). LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways (PubMed:21455173, PubMed:21455180, PubMed:21455181). Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation (PubMed:21455173, PubMed:21455180, PubMed:21455181). LUBAC is recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex (PubMed:21455173, PubMed:21455180, PubMed:21455181). The LUBAC complex is also involved in innate immunity by conjugating linear polyubiquitin chains at the surface of bacteria invading the cytosol to form the ubiquitin coat surrounding bacteria (PubMed:28481331). LUBAC is not able to initiate formation of the bacterial ubiquitin coat, and can only promote formation of linear polyubiquitins on pre-existing ubiquitin (PubMed:28481331). The bacterial ubiquitin coat acts as an 'eat-me' signal for xenophagy and promotes NF-kappa-B activation (PubMed:28481331). Together with OTULIN, the LUBAC complex regulates the canonical Wnt signaling during angiogenesis (PubMed:23708998).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal SHARPIN antibody. Suitable for WB and reacts with Human, Mouse samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR29120-74

Purification technique

Affinity purification Protein A

Specificity

Unsuitable for WB in human tissues.

Concentration
Loading...

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.

Supplementary info

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

Activity summary

SHARPIN also known as SHANK-associated RH domain-interacting protein is a multi-functional protein with a molecular mass of approximately 34 kDa. It primarily acts as a ubiquitin-binding protein and is a part of the LUBAC (Linear Ubiquitin Chain Assembly Complex) involved in various cellular signaling pathways. SHARPIN is expressed in a range of tissues but is highly abundant in lymphoid tissues and the brain. Its mechanical actions involve binding and modulating ubiquitin chains which plays a role in regulating protein stability and signaling processes in cells.

Biological function summary

SHARPIN engages in significant cellular functions by being a critical component of the LUBAC complex. This complex is central to the activation of NF-kB signaling by mediating linear ubiquitination of specific substrate proteins influencing inflammatory responses and cell death regulation. In the immune system SHARPIN regulates the TNF receptor signaling and helps maintain homeostasis. Its absence or mutation can lead to dysregulated cell survival and immune responses highlighting its importance in maintaining cellular functions.

Pathways

SHARPIN integrates into the NF-kB and Wnt signaling pathways which are vital for regulating immune responses and cell fate decisions. Within the NF-kB pathway SHARPIN cooperates with other LUBAC components such as HOIP and HOIL-1L to facilitate ubiquitination necessary for pathway activation. Additionally SHARPIN influences the Wnt pathway through interactions with Dishevelled proteins contributing to developmental and stem cell signaling. The coordination of these pathways by SHARPIN highlights its role in cellular communication and signaling.

Associated diseases and disorders

SHARPIN has connections to conditions such as chronic inflammation and immunodeficiency syndromes. Dysregulation in SHARPIN function can contribute to chronic inflammatory diseases including psoriasis due to improper NF-kB pathway regulation. Its association with HOIP and HOIL-1L through the LUBAC complex also links it to autoinflammatory conditions characterized by immune system imbalances. SHARPIN mutations are implicated in skin and immune disorders reflecting its broad impact on health and disease manifestation.

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.

3 product images

  • Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891), expandable thumbnail

    Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 32165044).

    The identity of the lower MW band at approximately 30 kDa (in lanes 1, 3 and 5) is unknown.

    Exposure time: Lanes 1-5: 147 seconds; Lane 6: 8 seconds

    All lanes: Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

    Lane 1: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

    Lane 2: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 6: CT26.WT (mouse colon fibroblast) 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 1/20000 dilution

    Observed band size: 40 kDa

  • Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891), expandable thumbnail

    Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 32165044).

    To minimize protein degradation, cells (lanes 7-9) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

    All lanes: Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

    Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 3: NIH/3T3 (mouse embryonic fibroblast) 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 1/20000 dilution

    Observed band size: 40 kDa

    Exposure time: 180s

  • Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891), expandable thumbnail

    Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 32165044).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

    All lanes: Western blot - Anti-SHARPIN antibody [EPR29120-74] (ab321891) at 1/1000 dilution

    Lane 1: THP-1 (human monocytic leukemia monocyte) transfected with scrambled siRNA control whole cell lysate at 20 µg

    Lane 2: THP-1 transfected with siRNA specifically targeting NTHP-1 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

    Developed using the ECL technique.

    Observed band size: 40 kDa, 36 kDa

    Exposure time: 37s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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