Cytokine receptor-like factor 3 is a protein that in humans is encoded by the CRLF3 gene. Model organisms have been used in the study of CRLF3 function. A conditional knockout mouse line, called Crlf3tm1a(KOMP)Wtsi was generated as part of the International Knockout Mouse Consortium program, a high-throughput mutagenesis project to generate and distribute animal models of disease to interested scientists. Male and female animals underwent a standardized phenotypic screen to determine the effects of deletion. Twenty six tests were carried out and two significant phenotypes were reported. Homozygous mutant female adults had a significant increase in circulating levels of fructosamine, while mutants of both sexes had decreased platelet cell numbers. Although CRLF3 signaling pathways have not yet been fully characterized it is very likely that CRLF3 is a neuroprotective erythropoietin receptor.
Background References
1. Taznin T et al. Cytokine Receptor-Like Factor 3 (CRLF3) Contributes to Early Zebrafish Hematopoiesis. Front Immunol. 2022 Jun
2. Bennett C et al. CRLF3 plays a key role in the final stage of platelet genesis and is a potential therapeutic target for thrombocythemia. Blood. 2022 Apr
Western blot analysis of CRLF3 on different lysates with Rabbit anti-CRLF3 antibody (HA722054) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: Raji cell lysate (20 µg/Lane) Lane 3: Ramos cell lysate (20 µg/Lane) Lane 4: K-562 cell lysate (20 µg/Lane) Lane 5: Jurkat cell lysate (20 µg/Lane) Lane 6: 293T cell lysate (20 µg/Lane) Lane 7: A549 cell lysate (20 µg/Lane) Lane 8: RAW264.7 cell lysate (20 µg/Lane) Lane 9: PC-12 cell lysate (20 µg/Lane) Lane 10: Mouse thymus tissue lysate (40 µg/Lane) Lane 11: Mouse spleen tissue lysate (40 µg/Lane) Lane 12: Rat thymus tissue lysate (40 µg/Lane) Lane 13: Rat testis tissue lysate (40 µg/Lane)
Predicted band size: 50 kDa Observed band size: 50 kDa Exposure time: 30 seconds; ECL: K1801; 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 (HA722054) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of RAW264.7 cells labeling CRLF3 with Rabbit anti-CRLF3 antibody (HA722054) 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-CRLF3 antibody (HA722054) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"