Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial cells)
Gel Running Conditions
Reduced Denaturing (12% acrylamide gel)
Loading amount
15 µg
Specification
Epithelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk
Dilution
1/1000
Secondary antibody
Dilution
1/40000
Name
Non-Abcam antibody was used: Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) (
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
None
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 35-50 kDa Non-specific: 15 kDa
Positive control
Image included in this review shows CAR staining with this product on transformed human intestinal epithelial cell lines (T84, SKCO-15, and Caco-2 BBE) which have been previously reported to express CAR. Calnexin is shown as a loading control.
Additional data
Additional Notes
Great antibody for western blot and immunofluorescence. Works in both BSA and milk blocking solutions and is cross reactive for murine primary intestinal epithelial cells (colon and ileum). Exhibits CAR specific staining by western blotting when using negative controls (CHO cells and primary murine bone marrow leukocytes).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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提交于 Nov 30 2021