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- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- MA5-16420 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin 8 Monoclonal Antibody (SP102)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is breast carcinoma.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SP102
- Vial size
- 500 µL
- Concentration
- 0.01 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references A small molecule inhibitor of SRC family kinases promotes simple epithelial differentiation of human pluripotent stem cells.
Genetic deletion of caspase-2 accelerates MMTV/c-neu-driven mammary carcinogenesis in mice.
Lian X, Selekman J, Bao X, Hsiao C, Zhu K, Palecek SP
PloS one 2013;8(3):e60016
PloS one 2013;8(3):e60016
Genetic deletion of caspase-2 accelerates MMTV/c-neu-driven mammary carcinogenesis in mice.
Parsons MJ, McCormick L, Janke L, Howard A, Bouchier-Hayes L, Green DR
Cell death and differentiation 2013 Sep;20(9):1174-82
Cell death and differentiation 2013 Sep;20(9):1174-82
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing ofCytokeratin 8 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR745510_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of Cytokeratin 8 was performed by loading 50 µg of HeLa Cas9 (Lane 1) and HeLa Cas9 cells transduced with Cytokeratin 8 Lentiviral sgRNA (Lane 2) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Cytokeratin 8 Monoclonal Antibody (SP102) (Product # MA5-16420) using 1:500 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5000 dilution).Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toCytokeratin 8 (Fig (b)).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Cytokeratin 8 was achieved by transfecting Caco-2 with Cytokeratin 8 specific siRNAs (Silencer® select Product # S7969, S7970). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Cytokeratin 8 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cytokeratin 8 Monoclonal Antibody (SP102) (Product # MA5-16420, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Cytokeratin 8.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cytokeratin 8 Monoclonal Antibody (SP102)(Product # MA5-16420) and a 53,57kDa band corresponding to Cytokeratin 8 was observed across the cell lines tested except IMR-32, SH-SY5Y and SK-MEL-31. Whole cell extracts (30 µg lysate) of Caco-2 (Lane 1), HaCaT (Lane 2), MCF7 (Lane 3), IMR-32 (Lane 4), SH-SY5Y (Lane 5) and SK-MEL-31 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 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 Cytokeratin 8 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with ice-cold acetone at 4°C for 5 minutes and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cytokeratin 8 Monoclonal Antibody (SP102) (Product # MA5-16420) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (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 cytoskeletal 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 Keratin 8 using anti-Keratin 8 Monoclonal Antibody (Product # MA5-16420) in Breast Ductal Cell Carcinoma Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.
