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ELISAAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- MA5-15357 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FER Monoclonal Antibody (5D2C4)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15357 targets FER in indirect ELISA, IF, IHC, and WB applications and shows reactivity with Human and mouse samples. The MA5-15357 immunogen is purified recombinant fragment of human FER expressed in E. Coli. MA5-15357 detects FER which has a predicted molecular weight of approximately 95kDa.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 5D2C4
- Vial size
- 100 µL
- Concentration
- Conc. Not Determined
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of FER using FER monoclonal antibody (Product # MA5-15357) in NIH/3T3 (1), A549 (2) and SK-MEL-5 (3) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of FER was achieved by transfecting HeLa with FER specific siRNAs (Silencer® select Product # s5109, s5111). Western blot analysis (Fig. a) was performed using whole cell extracts from the FER knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with FER Monoclonal Antibody (5D2C4) (Product # MA5-15357, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to FER.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), A549 (Lane 2), 3T3-L1 (Lane 3), HeLa (Lane 4), HT-1080 (Lane 5), SW-480 (Lane 6) and tissue extracts of Mouse Brain (Lane 7) and Rat Brain (Lane 8). The blot was probed with Anti-FER Monoclonal Antibody (5D2C4) (Product # MA5-15357, 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). A 95 kDa band corresponding to FER was observed across all cell lines and tissue extracts tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HeLa cells using FER monoclonal antibody (Product # MA5-15357) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of FER was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with FER Mouse Monoclonal Antibody (5D2C4) (Product # MA5-15357) 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, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human kidney tissues using FER monoclonal antibody (Product # MA5-15357).
