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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [3]
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- Product number
- MA5-27398 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CCDC22 Monoclonal Antibody (OTI2E4)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI2E4
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-CCDC22 Monoclonal Antibody (OTI2E4) (Product # MA5-27398) and a 70 kDa band corresponding to Coiled-coil domain-containing protein 22 was observed across all the tested cell lines. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), NIH:OVCAR-3 (Lane 2), SK-O-V3 (Lane 3), U-2 OS (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Coiled-coil domain-containing protein 22 was performed using 70% confluent log phase NIH:OVCAR-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CCDC22 Monoclonal Antibody (OTI2E4) (Product # MA5-27398) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), 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 dilution). 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
- Immunohistochemistry was performed on paraffin-embedded human prostate tissue. To expose target proteins, heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100°C for 10min. Following antigen retrieval, tissues were probed with a CCDC22 monoclonal antibody (Product # MA5-27398) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human bladder tissue. To expose target proteins, heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100°C for 10min. Following antigen retrieval, tissues were probed with a CCDC22 monoclonal antibody (Product # MA5-27398) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human tonsil tissue. To expose target proteins, heat-induced epitope retrieval by 10mM citric buffer, pH6.0, 100°C for 10min. Following antigen retrieval, tissues were probed with a CCDC22 monoclonal antibody (Product # MA5-27398) at a dilution of 1:500.
