Calnexin/CANX Mouse Monoclonal Antibody
| Cat Number: | MAB-94756 |
|---|---|
| Conjugate: | Unconjugated |
| Size: | 200 ug |
| Clone: | 8D10B3 |
| Concentration: | Adding 0.2 ml of distilled water will yield a concentration of 1mg/ml. |
| Host: | Mouse |
| Isotype: | IgG2b |
| Immunogen: | E.coli-derived human Calnexin/CANX recombinant protein. |
| Reactivity: | Human, Mouse, Rat |
| Applications: | Western blot: 1:1000 Immunocytochemistry: 1:100 Immunofluorescence: 1:100 |
| Molecular Weight: | 95 kDa |
| Purification: | Immunogen affinity purified. |
| Background: | Calnexin (CNX) is a 67 kDa integral protein of the endoplasmic reticulum. This gene encodes a member of the calnexin family of molecular chaperones. The encoded protein is a calcium-binding, endoplasmic reticulum (ER)-associated protein that interacts transiently with newly synthesized N-linked glycoproteins, facilitating protein folding and assembly. It may also play a central role in the quality control of protein folding by retaining incorrectly folded protein subunits within the ER for degradation. Alternatively spliced transcript variants encoding different isoforms have been described. |
| Form: | Lyophilized |
| Buffer: | Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4. |
| Storage: | At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing. |
Western blot analysis of Calnexin/CANX using anti- Calnexin/CANX antibody.
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 Jurkat whole cell lysates, Lane Lane 2: human placenta tissue lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: 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 mouse anti-
Calnexin/CANX antigen affinity purified monoclonal antibody at 1:1000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Calnexin/CANX at approximately 95 kDa. The expected band size for Calnexin/CANX is at 67 kDa.
HC analysis of Calnexin/CANX using anti- Calnexin/CANX antibody.
Calnexin/CANX was detected in a paraffin-embedded section of human endometrial adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:100 mouse anti-Calnexin/CANX Antibody overnight at 4°C.
Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was
developed using HRP Conjugated Mouse IgG Super VisionAssay Kit with DAB as the chromogen.
IHC analysis of Calnexin/CANX using anti- Calnexin/CANX antibody.
Calnexin/CANX was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen
retrieval was performed in EDTA buffer (pH 8.0, epitoperetrieval solution). The tissue section was blocked with 10%
goat serum. The tissue section was then incubated with 1:100 mouse anti-Calnexin/CANX Antibody.
overnight at 4°C. Peroxidase Conjugated Goat Anti-mouseIgG was used as secondary antibody and incubated for 30minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.
IF analysis of Calnexin/CANX using anti- Calnexin/CANX antibody.
Calnexin/CANX was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was
performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat
serum. And then incubated with 1:100 mouse anti- Calnexin/CANX Antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of A549 cells using anti-Calnexin/CANX antibody.
Overlay histogram showing A549 cells stained with (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- Calnexin/CANX Antibody 1 ug/1×106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG
( 5-10 ug/1×106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 ug/1×106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.