Calnexin Antibody - BSA Free

Western Blot: Calnexin Antibody [NB100-1974] - Identification of plasma exosomes and differentially expressed exosomal miRNAs. Confirmation of the exosomes markers with Western blotting indicated the presence of CD63 but the absence of calnexin in exosomes. Image collected and cropped by CiteAb from the following publication (https://www.frontiersin.org/article/10.3389/fonc.2019.00459/full), licensed under a CC-BY license.

Immunohistochemistry: Calnexin Antibody [NB100-1974] - Analysis of Calnexin in mouse bladder using DAB with hematoxylin counterstain.

Flow Cytometry: Calnexin Antibody [NB100-1974] - An intracellular stain was performed on rat FR cells with Calnexin Antibody NB100-1974 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: Calnexin Antibody [NB100-1974] - Analysis of Calnexin in HeLa whole cell lysate.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1974] - Zebrafish Vwf forms pseudo-Weibel-Palade bodies (pseudo-WPBs) in mammalian cell culture. pzVwf/Myc-HIS (zebrafish Vwf) plasmids were transfected into HEK293T cells. Anti-Myc antibody conjugated to Alexa Fluor 488 (green channel) was used for detection and anti-calnexin antibody conjugated to Alexa Fluor 594 (red channel) labeled endoplasmic reticulum (ER). Both constructs demonstrate formation of elongated Myc positive and ER negative structures (absence of yellow signal in the merged panels) characteristic of pseudo-WPBs. Scale bars, 2.5 um. Image collected and cropped by CiteAb from the following publication (https://www.hindawi.com/journals/ah/2012/214209/), licensed under a CC-BY license.

Flow Cytometry: Calnexin Antibody [NB100-1974] - Analysis of Alexa Fluor (R) 488 conjugate of NB100-1974. An intracellular stain was performed on Jurkat cells with Calnexin antibody NB100-1974AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange).

Flow Cytometry: Calnexin Antibody [NB100-1974] - An intracellular stain was performed on HeLa cells with Calnexin antibody NB100-1974AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488. Image using the Alexa Fluor 488 form of this antibody.

Flow Cytometry: Calnexin Antibody [NB100-1974] - An intracellular stain was performed on Daudi cells with Calnexin Antibody NB100-1974 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Flow Cytometry: Calnexin Antibody [NB100-1974] - An intracellular stain was performed on NIH3T3 cells with Calnexin Antibody NB100-1974 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Simple Western: Calnexin Antibody [NB100-1974] - Image shows a specific band for Calnexin in 0.1 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: Calnexin Antibody - BSA Free [NB100-1974] -

Identification of plasma exosomes and differentially expressed exosomal miRNAs. (D) Confirmation of the exosomes markers with Western blotting indicated the presence of CD63 but the absence of calnexin in exosomes. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31249805), licensed under a CC-BY licence.

Western Blot: Calnexin Antibody - BSA Free [NB100-1974] -

EMPA treatment attenuated cardiomyocyte overproduction of reactive oxygen species (ROS) induced by TAC. (A) Left, Mitosox staining of cardiomyocytes in sham, sham + EMPA, TAC and TAC + EMPA groups and Right, quantitative results. (B) Representative blots of NRF-2 and HO-1, and quantitative results. (C) The relative mRNA levels of genes related to endogenous antioxidants. Results are expressed as mean ± SEM, n = 5–7, *p 0.05 vs. corresponding sham group, †p 0.05 vs. corresponding TAC vehicle group. One-way ANOVA and Tukey post hoc test. EMPA, empagliflozin; SEM, standard error of the mean; TAC, transverse aortic constriction; NRF-2, the nuclear factor erythroid 2–related factor 2; HO-1, heme oxygenase-1; GCLM, glutamate-cysteine ligase modifier subunit; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35647080), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Calnexin Antibody - BSA Free [NB100-1974] -

HIF‐2 alpha depletion counteracts tumour growth and non‐canonical glutamine metabolism in vivo. (A) Representative images of tumour‐bearing mice and tumours removed from the mice. (B) Tumour volumes were measured on the indicated days. (C) Tumour weights were shown as the means ± S.D. when the tumours were harvested. (D) Representative images (×200) of H&E and IHC staining of the tumours. Ki67 was used as proliferation index. (E and F) qRT‐PCR and Western blot analysed the expression of HIF‐2 alpha and GOT1 in tumour tissues from sh‐HIF‐2 alpha Panc‐1 group compared with control group, *P 0.05, **P 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28544376), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Calnexin Antibody - BSA Free [NB100-1974] -

Bmal1 regulates tumor-associated macrophage polarization.(A) Bmal1 gene expression (left panel) and protein levels (right panel) in WT macrophages treated with control medium or increasing doses of B16-F10 tumor-conditioned medium (T-CM, diluted 1:3 or 1:1 with control medium) for 8 hr. N = 3 biological replicates for qPCR, statistical analysis was performed using Student’s T test. (B) Measurement of mROS using MitoSox Red (mean fluorescent intensity, MFI) in mitochondria from macrophages treated with control medium or T-CM diluted 1:1 with control medium for 1 hr. N = 3 biological replicates, statistical analysis was performed using Student’s T test. (C) Hif-1 alpha protein levels in WT and M-BKO macrophages treated with control medium, T-CM diluted 1:1 with control medium, or undiluted T-CM for 4 hr. (D) Glycolytic stress test in macrophages pretreated with control medium or T-CM diluted 1:1 with control medium for 4 hr. N = 5 biological replicates. Statistical analysis was performed using two-way ANOVA comparing T-CM-treated M-BKO with WT cells across the time course. (E) Relative expression of genes involved in amino-acid metabolism and oxidative stress response in macrophages treated with control medium or T-CM diluted 1:3 with control medium for 8 hr, as determined by qPCR. N = 3 biological replicates, statistical analysis was performed using Student’s T test. (F) Tumor volume in male (left) and female (right) WT and M-BKO mice. 300,000 B16-F10 cells were injected subcutaneously into the right flank. N = 18 (male) and 8 (female) mice, statistical analysis was performed using two-way ANOVA for WT vs M-BKO mice across the time course. (G) Gene expression for F4/80+ cells isolated from B16-F10 tumors or spleens of female mice 14 days after injection. Tissues from six mice per genotype were pooled into three groups for leukocyte isolation. Statistical analysis was performed using Student’s T test. Data are presented as mean ± S.E.M. *, p0.05. Experiments were repeated at least twice.Increased mROS levels and glucose uptake in M-BKO TAMs.(A) Tumor volumes of B16-F10 subcutaneous allografts in 3-month-old female WT and M-BKO mice 14 days after tumor cell injection. Each mouse was injected with 500,000 B16-F10 cells in the right and left flanks, and tumors from each mouse were pooled for immune cell isolation and flow cytometry. N = 4 mice, with two tumors per mouse. (B, C) mROS production and glucose uptake by tumor-associated CD45+F4/80+ cells determined by flow cytometry (mean fluorescent intensities) of Mitosox Red and 2-NBDG staining, respectively. N = 4. Statistical analysis was performed using Student’s T test. Data are presented as mean ± S.E.M. *, p0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32396064), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Calnexin Antibody - BSA Free [NB100-1974] -

Tau clearance in vivo and autophagy function. Clearance kinetics of photoconverted Dendra-tau measured in neurons of WT-tau and AT152T-tau fish. Measurement of the intensity of the red Dendra-tau signal over time reflects the clearance or degradation of tau protein. (A) Representative images of photoconverted red Dendra-tau signal comparing a single neuron from WT-tau and A152T-tau fish at three different timepoints: immediately after photoconversion (0 h), 24 h and 48 h after photoconversion. (B) Quantification of red Dendra-tau intensity in photoconverted neurons in the spinal cord of WT-tau and A152T-Tau transgenic fish (representative images shown in Supplementary Fig. 8C). The percentage of residual photoconverted red Dendra-tau was measured over 48 h measured at 12-h intervals. Dendra-tagged A152T-tau clears at a significantly lower rate than WT-tau. (n = 30/group shown as mean ± SD; Student-Newman-Keuls one-way ANOVA, **P 0.01 and ***P 0.001 versus WT-tau). (C–F) Western blots for LC3-II, a well-characterized marker of autophagosome number, demonstrate that there are no differences in the levels of this protein between WT-tau and A152T-tau fish either at 24 hpf (pre-phenotype; C and D) or 72 hpf (post-phenotype; E and F). (E and F) Measurements of LC3-II levels in the presence or absence of ammonium chloride provides a method for measuring autophagic flux. No differences were observed between the two transgenic lines at 3 dpf, suggesting that autophagy functions normally in both WT-tau and A152T-tau fish (graph represents mean ± SD of four independent clutches per group for E and F and three for C and D; two-tailed t-test). (G and H) Clearance kinetics of Dendra-tau was measured in the presence or absence of ammonium chloride. Treatment with ammonium chloride blocks autophagic flux and delays the clearance of both WT-tau and A152T-tau to the same extent, indicating that flux occurs at the same rate in these two lines (mean ± SD, n = 62 neurons/group; Student-Newman-Keuls one-way ANOVA, **P 0.01 and ***P 0.001 versus untreated group). Note in G and H, the ‘WT-tau + NH4Cl’ (denoted by black squares and black dashed line) overlaps with the ‘A152T-tau’ line (denoted by grey triangles and grey solid line). The graphs in G and H are presented with a different line in the foreground and background to aid interpretation. Image collected and cropped by CiteAb from the following open publication (https://academic.oup.com/brain/article/140/4/1128/2980948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Calnexin in U-2 OS Human Cell Line.

Calnexin was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-Calnexin Affinity Purified Polyclonal Antibody conjugated to FITC (Catalog # NB100-1974F) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.