WB
Western blot analysis of TRIM21 on different lysates with Rabbit anti-TRIM21 antibody at 1/10,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 10ng/mL IFN-α for 16 hours cell lysate, Lane 3: A549 cell lysate, Lane 4: 293T cell lysate, Lane 5: U-2 OS cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 30 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-TRIM21 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-TRIM21 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of U-2 OS cells labeling TRIM21 with Rabbit anti-TRIM21 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TRIM21 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of U-2 OS cells labeling TRIM21. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
TRIM21 was immunoprecipitated from 0.2 mg A549 cell lysate with Rabbit anti-TRIM21 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-TRIM21 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input), Lane 2: Rabbit anti-TRIM21 antibody IP in A549 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-TRIM21 antibody in A549 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 6 seconds.| Product Name | TRIM21 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human TRIM21 aa 1-475 (P19474). |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000-1:10000 IHC:1:200-1:1000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | TRIM21 |
|---|---|
| Gene Synonyms | SSA RO52 SSA1 RNF81 Ro/SSA |
| Gene Full Name | tripartite motif containing 21 |
| Gene Summary | This gene encodes a member of the tripartite motif (TRIM) family. The TRIM motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region. The encoded protein is part of the RoSSA ribonucleoprotein, which includes a single polypeptide and one of four small RNA molecules. The RoSSA particle localizes to both the cytoplasm and the nucleus. RoSSA interacts with autoantigens in patients with Sjogren syndrome and systemic lupus erythematosus. Alternatively spliced transcript variants for this gene have been described but the full-length nature of only one has been determined. [provided by RefSeq, Jul 2008] |
| Alternative Names | 52 kDa ribonucleoprotein autoantigen Ro/SS-A antibody 52 kDa Ro protein antibody 52kD Ro/SSA autoantigen antibody Autoantigen Ro/SSA 52-KD antibody E3 ubiquitin-protein ligase TRIM21 antibody RING finger protein 81 antibody RNF81 antibody Ro 52 antibody Ro(SS-A) antibody Ro52 antibody 52 kDa ribonucleoprotein autoantigen Ro/SS-A antibody 52 kDa Ro protein antibody 52kD Ro/SSA autoantigen antibody Autoantigen Ro/SSA 52-KD antibody E3 ubiquitin-protein ligase TRIM21 antibody RING finger protein 81 antibody RNF81 antibody Ro 52 antibody Ro(SS-A) antibody Ro52 antibody RO52_HUMAN antibody Sicca syndrome antigen A antibody Sjoegren syndrome type A antigen antibody Sjogren syndrome antigen A1 antibody Sjogren syndrome type A antigen antibody SS-A antibody SSA antibody SSA1: Sjogren syndrome antigen A1 (52kDa ribonucleoprotein autoantigen SS-A/Ro) antibody TRIM21 antibody Tripartite motif protein TRIM21 antibody Tripartite motif-containing 21 antibody Tripartite motif-containing protein 21 antibody |
| Molecular Weight(MW) | 54kDa(Observed band size: 50 kDa) |
| Cellular Localization | Cytoplasm, Cytoplasmic vesicle, autophagosome, Nucleus, P-body. |
WB
Western blot analysis of TRIM21 on different lysates with Rabbit anti-TRIM21 antibody at 1/10,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 10ng/mL IFN-α for 16 hours cell lysate, Lane 3: A549 cell lysate, Lane 4: 293T cell lysate, Lane 5: U-2 OS cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 30 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-TRIM21 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-TRIM21 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of U-2 OS cells labeling TRIM21 with Rabbit anti-TRIM21 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TRIM21 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of U-2 OS cells labeling TRIM21. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
TRIM21 was immunoprecipitated from 0.2 mg A549 cell lysate with Rabbit anti-TRIM21 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-TRIM21 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input), Lane 2: Rabbit anti-TRIM21 antibody IP in A549 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-TRIM21 antibody in A549 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 6 seconds.| Application Notes | WB:1:5000-1:10000 IHC:1:200-1:1000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
