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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- MA1-91326 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin Pan Monoclonal Antibody (B311.1)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- High temperature antigenic unmasking technique or enzymatic predigestion is recommended with this antibody when using paraffin-embedded tissue sections. Positive control of skin or colonic carcinoma suggested. This antibody detects Cytokeratin 10, Cytokeratin 13, Cytokeratin 18, Cytokeratin 4, Cytokeratin 5, Cytokeratin 6 and Cytokeratin 8.
- Reactivity
- Human, Mouse, Rat, Bovine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- B311.1
- Vial size
- 500 µL
- Concentration
- 0.4 mg/mL
- Storage
- 4° C, do not freeze
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), MCF7 (Lane 2), HeLa (Lane 3), Caco-2 (Lane 4), SW-480 (Lane 5), Hep G2 (Lane 6), T-47D (Lane 7), NIH/3T3 (Lane 8), MOLT-4 (Lane 9) and Raji (Lane 10). The blot was probed with Cytokeratin Pan Monoclonal Antibody (Product # MA1-91326, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). Bands between 40 to 56 kDa were observed across cell lines (except MOLT-4 and Raji which are reported to be negative) which corresponds to different type of cytokeratins.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Cytokeratin Pan was performed using 70% confluent log phase A-431 cells. The cells were fixed and permeabilized with ice-cold 100% acetone and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin Pan Monoclonal Antibody (B311.1) (Product # MA1-91326) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing cytoskeletal localization. The images were captured at 60X magnification.
