Knockout Tested Rabbit Polyclonal RNF31/HOIP antibody. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 24 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
WB | ICC/IF | |
---|---|---|
Human | Tested | Tested |
Mouse | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
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E3 ubiquitin-protein ligase component of the LUBAC complex which conjugates linear ('Met-1'-linked) polyubiquitin chains to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation (PubMed:17006537, PubMed:19136968, PubMed:20005846, PubMed:21455173, PubMed:21455180, PubMed:21455181, PubMed:22863777, PubMed:28189684, PubMed:28481331). 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:17006537, PubMed:19136968, PubMed:20005846, PubMed:21455173, PubMed:21455180, PubMed:21455181, PubMed:22863777, PubMed:28189684). Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation (PubMed:21455173, PubMed:28189684). 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:20005846, PubMed:27458237). 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, PubMed:34012115). 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). Recruited to the surface of bacteria by RNF213, which initiates the bacterial ubiquitin coat (PubMed:34012115). The bacterial ubiquitin coat acts as an 'eat-me' signal for xenophagy and promotes NF-kappa-B activation (PubMed:28481331, PubMed:34012115). Together with OTULIN, the LUBAC complex regulates the canonical Wnt signaling during angiogenesis (PubMed:23708998). RNF31 is required for linear ubiquitination of BCL10, thereby promoting TCR-induced NF-kappa-B activation (PubMed:27777308). Binds polyubiquitin of different linkage types (PubMed:23708998).
ZIBRA, RNF31, E3 ubiquitin-protein ligase RNF31, HOIL-1-interacting protein, RING finger protein 31, RING-type E3 ubiquitin transferase RNF31, Zinc in-between-RING-finger ubiquitin-associated domain protein, HOIP
Knockout Tested Rabbit Polyclonal RNF31/HOIP antibody. Suitable for WB, ICC/IF and reacts with Human samples. Cited in 24 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
Replenishment batches of ab46322 are tested in WB. Previous batches were additionally validated in ICC/IF. This application is still expected to work and is covered by our Abpromise guarantee.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
RNF31 also known as HOIP (HOIL-1 Interacting Protein) is a large protein with a molecular weight of around 119 kDa. It is a part of the ubiquitin-proteasome system and functions mechanically as a E3 ubiquitin ligase. RNF31 forms the catalytic component of the Linear Ubiquitin Chain Assembly Complex (LUBAC). Expression of RNF31 occurs in many tissues with notable levels in immune cells and the testes. The protein features a RING-in-between-RING (RBR) motif structure that is critical for its ligase activity.
RNF31/HOIP plays an important role in immune response regulation. It functions as part of the LUBAC complex alongside SHARPIN and RBCK1 (also known as HOIL-1L) which together catalyze the linear ubiquitination of substrates. These modifications regulate NF-kB signaling which is an important pathway involved in immune and inflammatory responses. RNF31's ubiquitin ligase activity adds linear ubiquitin chains to target proteins influencing their stability and function therefore impacting the immune cell function and survival.
RNF31/HOIP integrates into the NF-kB and the apoptosis pathways. In the NF-kB pathway it acts as a regulator of cytokine production and cell survival. RNF31 achieves this through modification of NEMO (NF-kB essential modulator) an important protein in this pathway. In apoptosis regulation linear ubiquitination by RNF31 modulates the activity of proteins such as caspases preventing inappropriate cell death during immune responses. This helps to maintain balance between survival and death signals in cells allowing for proper immune function.
RNF31/HOIP has implications in inflammatory diseases and certain cancers. Dysregulation of LUBAC components including HOIP has been linked with chronic inflammatory conditions like rheumatoid arthritis where inappropriate NF-kB activation occurs. Furthermore aberrations in RNF31 activity have associations with tumorigenesis where its altered signaling pathways can lead to uncontrolled cell proliferation. In these contexts proteins such as A20 may interact with RNF31 affecting the regulation of NF-kB signaling therefore influencing disease progression.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Western blot analysis of A549 cell lysate (100μg/lane) RNF31/HOIP with ab46322 at 1/1000. A HRP-conjugated Goat anti-rabbit polyclonal (1/10000) was used as the secondary antibody.
lane 1-2 : siCT A549
lane 3-4 : siHOIP A549
All lanes: Western blot - Anti-RNF31/HOIP antibody (ab46322)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 120 kDa
Exposure time: 2min
All lanes: Western blot - Anti-RNF31/HOIP antibody (ab46322) at 1/250 dilution
All lanes: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
All lanes: IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 120 kDa
Observed band size: 105 kDa, 116 kDa, 50 kDa, 60 kDa
ICC/IF image of ab46322 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46322, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293 cells at 5µg/ml.
Western blot: Anti-RNF31 antibody (ab46322) staining at 1/250 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab46322 was shown to bind specifically to RNF31. A band was observed at 130 kDa in wild-type A549 cell lysates with no signal observed at this size in RNF31 knockout cell line. To generate this image, wild-type and RNF31 knockout A549 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-RNF31/HOIP antibody (ab46322) at 1/250 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: RNF31 knockout A549 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
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.
Predicted band size: 120 kDa
Observed band size: 130 kDa
Western blot: Anti-RNF31 antibody (ab46322) staining at 1/250 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab46322 was shown to bind specifically to RNF31. A band was observed at 130-140 kDa in wild-type A549 cell lysates with no signal observed at this size in RNF31 knockout cell line. To generate this image, wild-type and RNF31 knockout A549 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-RNF31/HOIP antibody (ab46322) at 1/250 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: RNF31 knockout A549 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
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.
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