rabbit polyclonal anti-human calnexin antibodies  (ABclonal Biotechnology)

Journal: Biochemical and biophysical research communications

Article Title: Choline kinase alpha regulates autophagy-associated exosome release to promote glioma cell progression.

doi: 10.1016/j.bbrc.2024.151269

Figure Lengend Snippet: Fig. 3. CHKA mediates exosomes secretion in glioma cells. A-C Western blot (WB) analysis of exosome marker proteins CD63 and TSG101 in exosomes isolated from the supernatant of glioma cells in shCHKA and shNC group along with the negative control protein Calnexin. D Exosome protein concentration in the shCHKA and shNC groups of glioma cells using the BCA assay. E Electron microscopy was performed on shCHKA and shNC cells in glioma cells (Scale bars = 1 μm). F The number of multivesicular bodies (MVBs) per glioma cell profile. G The number of intraluminal vesicles (ILVs) per MVB profile in glioma cells. *P < 0.05, **P < 0.01, ***P < 0.001. shCHKA: CHKA knockdown group, shNC: con trol group.

Article Snippet: Antibodies used in the Western blotting process included: rabbit antihuman CHKA polyclonal antibody (abcam, USA), rabbit anti-human GAPDH polyclonal antibody (Affinity, China), rabbit anti-human CD63 polyclonal antibody (Proteintech, China), rabbit anti-human TSG101 polyclonal antibody(Proteintech, China), rabbit anti-human calnexin polyclonal antibody(Proteintech, China), rabbit anti-human LC3B polyclonal antibody(Sigma, USA), and rabbit anti-human P62/SQSTM1 polyclonal antibody(Sigma, USA).

Techniques: Western Blot, Marker, Isolation, Negative Control, Protein Concentration, BIA-KA, Electron Microscopy, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Comprehensive Analysis of Exosomal MicroRNAs Derived from UVB-Irradiated Keratinocytes as Potential Melanogenesis Regulators

doi: 10.3390/ijms25063095

Figure Lengend Snippet: Exosomes derived from UVB-irradiated keratinocytes are isolated and characterized by their size. ( A ) Confirmation of exosome specific markers such as CD9 and CD81 using Western blot. Negative exosomal marker calnexin was lost in the samples. ( B ) Exosome size distribution measured using electrophoretic trace. ( C ) Size distribution of microvesicles (nm) assessed using nanoparticle tracking analysis. The NTA results of keratinocyte cell-derived exosomes showed a 114 ± 6.5 nm mode size of the exosomes.

Article Snippet: The antibodies used were as follows: B-actin antibody (Sigma, St. Louis, MO, USA), MITF antibody (ThermoFisher Scientific, Fremont, CA, USA), rabbit anti-tyrosinase (Abcam, Cambridge, UK: ab170905), rabbit anti-TRP1 (Abcam, ab178676), rabbit anti-TRP2/DCT (Abcam, ab221144), anti-CD63 antibody (mouse monoclonal to CD63, ab59479), anti-human CD9 IgG rabbit polyclonal (System Biosciences, Palo Alto, CA, USA), and anti-human calnexin rabbit polyclonal (Cell signaling Technology, Danvers, MA, USA, Beverly; National Institute of Health, Bethesda, MD, USA).

Techniques: Derivative Assay, Irradiation, Isolation, Western Blot, Marker

Journal: Brain

Article Title: SLC30A9 mutation affecting intracellular zinc homeostasis causes a novel cerebro-renal syndrome

doi: 10.1093/brain/awx013

Figure Lengend Snippet: SLC30A9 protein subcellular localization, effect on Wnt signaling and structural modeling. ( A ) Overexpression of SLC30A9 protein fused to enhanced green fluorescent protein (EGFP) and immunofluorescent staining of different endosomal, Golgi and ER markers (CD63, EEA1, Rab7, GM130 and Calnexin) showing partial co-localization with endoplasmic reticulum in SH-SY5Y cells. On the bottom left side of the merged image of Calnexin marker panel is a magnification of the dashed frame, showing the co-localization in a single cell. Scale bars = 10 µm. ( B ) TOP flash reporter assay of neuroblastoma (SH-SY5Y) cells showing enhanced Wnt signalling in both SLC30A9 wild-type and mutant transfected cells. There was no significant difference between wild-type and mutant transfected cells. ( C ) Structural model of ZnT-9 protein showing the p.(A350del) in the fourth transmembrane helix, putatively causing destabilization of the protein structure in general, affecting the TM metal binding site specifically.

Article Snippet: Cells were then incubated with primary polyclonal goat anti-human EEA1 (Sc-6414; Santa Cruz Biotechnology), polyclonal rabbit anti- human Rab7 (Sc-10767; Santa Cruz Biotechnology), mouse monoclonal anti- human CD63 (CBL553; Merck Millipore) or polyclonal rabbit anti-human Calnexin (Sc-11397; Santa Cruz Biotechnology) antibodies for 1 h. Post-incubation, cells were washed twice with PBST and incubated with secondary donkey anti-goat IgG Alexa Fluor® 546 (A-11056; Invitrogen), goat anti-rabbit IgG Alexa Fluor® 546 (A-11010; Invitrogen) or donkey anti-mouse IgG Alexa Fluor® 546 (A-10036; Invitrogen) accordingly for 1 h. Cells were then washed twice with 1 × PBST and mounted using Vectashield containing DAPI (H-1200; Vector Laboratories).

Techniques: Over Expression, Staining, Marker, Reporter Assay, Mutagenesis, Transfection, Binding Assay