Recombinant Mouse CARD11 Protein

• Cat.No.: CARD11-2728M
• Product Overview: Recombinant Mouse CARD11 full length or partial length protein was expressed.

Specification

• Species: Mouse
• Source: Mammalian Cells
• Tag: His
• Form: Liquid or lyophilized powder
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: >80%
• Notes: This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
• Storage: Store it at +4 ºC for short term. For long term storage, store it at -20 ºC～-80 ºC.
• Storage Buffer: PBS buffer

Gene Information

• Gene Name: Card11 caspase recruitment domain family, member 11 [ Mus musculus ]
• Official Symbol: CARD11
• Gene ID: 108723
• mRNA Refseq: NM_175362.2
• Protein Refseq: NP_780571.2
• UniProt ID: Q8CIS0

Citation

TRIM41 is required to innate antiviral response by polyubiquitinating BCL10 and recruiting NEMO

Journal: Signal Transduction and Targeted Therapy    PubMed ID: 33640899    Data: 2021/2/28

Authors: Zhou Yu, Xuelian Li, Xuetao Cao

Article Snippet:Recombinant CARD9 (ab131800) and BCL10 (ab82241) were obtained from Abcam Inc. (Cambridge, MA).Recombinant CARD9 (ab131800) and BCL10 (ab82241) were obtained from Abcam Inc. (Cambridge, MA).. Recombinant CARD11 was obtained from Creative BioMart (Shirley, NY; Card11-385M).. Other non-specified reagents were purchased from Sigma-Aldrich (St. Louis, MO).Other non-specified reagents were purchased from Sigma-Aldrich (St. Louis, MO).

The PRY-SPRY domain of TRIM41 directly binds the CARD domain of BCL10. a Wild type BMDM were infected with VSV (MOI = 1) or HSV-1 (MOI = 5) viruses for 2 h as indicated. Then whole cell extracts (WCE) were immunoprecipitated with anti-TRIM41 antibody plus protein A/G beads. Components in the TRIM41 complex were examined by Western blotting. b 1 μg GST or GST-TRIM41 was coincubated with 1 μg recombinant CARD9, CARD11, or BCL10 for 1 h. Then sepharose 4B beads were used to pull down the GST complex. Components pulled down by the beads were examined by Western blotting. c Schematic illustration of domains in full-length (FL) or fragments (F) of TRIM41-Flag or BCL10-Myc. R, ring finger domain; B, B-box domain; CC, coiled-coil domain; CTD, C-terminal domain. Numerical numbers indicate the site of amino acids. d , e HEK293T cells were transiently transfected with Flag-tagged TRIM41 (or mutants) and Myc-tagged BCL10 (or mutants) as indicated for 48 h. Whole cell extracts (WCE) were immunoprecipitated with anti-Flag Sepharose Beads ( d ) or anti-Myc Sepharose Beads ( e ), and the associated BCL10 or TRIM41 was examined by Western blotting. f , g RAW264.7 cells grown on cover slides were transiently transfected with green fluorescent protein (GFP)-tagged TRIM41 and red fluorescent protein (RFP)-tagged BCL10 for 48 h, and then infected with VSV (MOI = 1) or HSV-1 (MOI = 5) viruses for 2 h. After counterstained with DAPI, cells were examined by confocal microscope ( f ; Scale bar, 100 nm). Ten double-positive randomly-selected cells on cover slides were measured for fluorescence colocalization, and the data are presented as mean ± SD ( g )

TRIM41 mediates Lys63-linked polyubiquitination of BCL10. a - c Trim41 +/+ or Trim41 –/– BMDM were infected with VSV (MOI = 1) or HSV-1 (MOI = 5) viruses for 2 h. Then whole-cell extracts (WCE) heated in buffer containing 1% SDS were immunoprecipitated (IP) with anti-BCL10 ( a ), anti-CARD9 ( b ), or anti-CARD11 ( c ) antibody plus protein A/G beads. Polyubiquitination of BCL10 ( a ), CARD9 ( b ), and CARD11 ( c ) was examined by Western blotting. d HEK293T cells were transiently transfected with Flag-tagged TRIM41 (or mutant) and Myc-tagged BCL10 vectors as indicated for 48 h. Then polyubiquitination of BCL10 was examined by Western blotting after immunoprecipitations with anti-Myc Sepharose Beads. e HEK293T cells were transiently transfected with Flag-tagged TRIM41, Myc-tagged BCL10, and HA-tagged Ub (mutants) vectors as indicated for 48 h. Then polyubiquitination of BCL10 was examined by Western blotting after immunoprecipitations with anti-Myc Sepharose Beads. f After incubation for 30 min, the in vitro polyubiquitination system was boiled for 5 min, and then polyubiquitinated BCL10 was examined by Western blotting against Ub after immunoprecipitations. g HEK293T cells were transiently transfected with Flag-tagged TRIM41 and Myc-tagged BCL10 (mutant) vectors as indicated for 48 h. Then polyubiquitination of BCL10 was examined by Western blotting after immunoprecipitations with anti-Myc Sepharose Beads. One representative experiment of three is shown