Anti-Calreticulin Rabbit Monoclonal Antibody

(A) CFRI-mediated ICD and DCs matured in vitro. Mechanism scheme of PTT-induced immunogenic death; 1,064-nm laser irradiation (1.0 W/cm 2 ) with or without treatment with DMEM, FR837, FIR1048, FRI, and CFRI (2 μg/ml). (B) The efflux of HMGB1 was quantitatively analyzed by flow cytometry. (C) CLSM images of CRT expression on the 4T1 cell surface after different treatments (scale bar: 100 μm). (D) Mean fluorescence intensity of CRT expression in 4T1 cells after different treatments. (E) The average fluorescence intensity of ATP exposure in 4T1 cells after different treatments. (F) The corresponding quantification (CD80 + CD86 + ) of mature DCs was quantitatively analyzed by flow cytometry. (G) After different treatments by enzyme-linked immunosorbent assay (ELISA), secretion levels of TNF-ɑ in the cell supernatant. (H) After different treatments by ELISA, the secretion level of IL-6 in the cell supernatant. (I) After different treatments by ELISA, the secretion level of IL-10 in the cell supernatant. Information of each group: I, control; II, control + L; III, FR837; IV, FR837 + L; V, FIR1048; VI, FIR1048 + L; VII, FRI; VII, FRI + L; IX, CFRI; X, CFRI + L. Significance is determined using one-way analysis of variance (* P < 0.05, ** P < 0.01, and *** P < 0.001). HSPs, heat shock proteins.
Index in PubMed under a CC BY license. PMID: 40040955

In vivo CFRI-mediated PTT-enhanced immunotherapy elicited an immune response after 16 d of treatment. (A) Schematic diagram of immunity induced by damage-associated molecular patterns (DAMPs). (B and C) Analysis of DC maturity (CD11 + CD80 + CD86 + ) using flow cytometry in the primary tumor of 4T1 tumor-bearing mice. (D and E) Regulatory T cells (Tregs) (CD4 + CD25 + ) in the spleen. (F and G) T cell proliferation in the blood (CD4 + CD8 + ). (H) The content of TNF-α in the spleen of 4T1 tumor-bearing mice. (I) The content of IL-6 in the spleen of 4T1 tumor-bearing mice. (J) The content of IL-10 in the spleen of 4T1 tumor-bearing mice. (K) Heat map of TNF-α and IL-10 content in the blood of 4T1 tumor-bearing mice. (L) CRT and HMGB1 staining images of 4T1 tumor-bearing mice after different treatments. Scale bar: 100 μm. Group information: I, PBS; II, PBS + L; III, FR837; IV, FR837 + L; V, FIR1048; VI, FIR1048 + L; VII, FRI; VIII, FRI + L; IX, CFRI; X, CFRI + L (* P < 0.05, ** P < 0.01, and *** P < 0.001). CTL, cytotoxic T lymphocyte; TCR, T cell receptor.
Index in PubMed under a CC BY license. PMID: 40040955

Western blot analysis of Calreticulin using anti-Calreticulin antibody (M00894).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human COLO320 whole cell lysates,
Lane 2: human HL-60 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat liver tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Calreticulin antigen affinity purified monoclonal antibody (Catalog # M00894) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Calreticulin at approximately 60 kDa. The expected band size for Calreticulin is at 48 kDa.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex, using the Antibody at 1:150 dilution.

Immunohistochemical analysis of paraffin-embedded Human renal cancer, using the Antibody at 1:150 dilution.

Immunohistochemical analysis of paraffin-embedded Human placenta, using the Antibody at 1:150 dilution.

Immunohistochemical analysis of paraffin-embedded Mouse testis, using the Antibody at 1:150 dilution.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum, using the Antibody at 1:150 dilution.

Immunofluorescent analysis using the Antibody at 1:50 dilution.