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Catalog Number | orb1728194 |
---|---|
Category | Antibodies |
Description | Anti-CD326/Epcam Antibody. Tested in ELISA, Flow Cytometry, IF, IHC, WB applications. This antibody reacts with Mouse, Rat. |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, IF, IHC, WB |
Reactivity | Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived mouse CD326/Epcam recombinant protein (Position: Q24-A267). |
Antibody Type | Primary Antibody |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 38 kDa |
UniProt ID | Q99JW5 |
Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
Alternative names | Agouti-related protein; Agrp; Agrt; Art Read more... |
Note | For research use only |
Application notes | Western blot, 0.1-0.25 μg/ml, Mouse Immunohistochemistry(Paraffin-embedded Section), 1-2 μg/ml, Mouse, Rat Immunofluorescence, 5 μg/ml, Mouse, Rat Flow Cytometry (Fixed), 1-3 μg /1x106 cells, Mouse ELISA, 0.1-0.5 μg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml |
Expiration Date | 12 months from date of receipt. |
IF analysis of CD326/Epcam using anti-CD326/Epcam antibody. CD326/Epcam was detected in a paraffin-embedded section of mouse colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL rabbit anti-CD326/Epcam Antibody overnight at 4°C. FITC Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 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.
IF analysis of CD326/Epcam using anti-CD326/Epcam antibody. CD326/Epcam was detected in a paraffin-embedded section of rat colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL rabbit anti-CD326/Epcam Antibody overnight at 4°C. FITC Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 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 RAW264.7 cells using anti-CD326/Epcam antibody. Overlay histogram showing RAW264.7 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD326/Epcam Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
IHC analysis of CD326/Epcam using anti-CD326/Epcam antibody. CD326/Epcam was detected in a paraffin-embedded section of mouse colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-CD326/Epcam Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
IHC analysis of CD326/Epcam using anti-CD326/Epcam antibody. CD326/Epcam was detected in a paraffin-embedded section of rat colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-CD326/Epcam Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Western blot analysis of CD326/Epcam using anti-CD326/Epcam 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: mouse small intestine tissue lysates, Lane 2: mouse kidney 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-CD326/Epcam antigen affinity purified polyclonal antibody at 0.25 µg/mL 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 with Tanon 5200 system. A specific band was detected for CD326/Epcam at approximately 38 kDa. The expected band size for CD326/Epcam is at 35, 40 kDa.
ELISA, FC, IF, IHC, WB | |
Mouse, Rat | |
Rabbit | |
Polyclonal | |
iFluor647 |