Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [3]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA5-29670 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin 5 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2. Predicted reactivity: Mouse (97%), Rat (97%), Dog (97%), Chicken (80%), Chimpanzee (100%), Bovine (97%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.35 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Cytokeratin 5 using 30 µg of HCT116 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Cytokeratin 5 polyclonal antibody (Product # PA5-29670) at a dilution of 1:5000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of Cytokeratin 5 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) at a dilution of 1:5000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Cytokeratin 5 was achieved by transfecting A-431 with Cytokeratin 5 specific siRNAs (Silencer® select Product # s7956, S7957). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Cytokeratin 5 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670, 1:5000 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 5.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) and a 60kDa band corresponding to Cytokeratin 5 was observed across the cell lines tested except MCF7, IMR-32 and SH-SY5Y along with non specific bands around 30-55 kDa. Whole cell extracts (30 µg lysate) of MCF 10A (Lane 1), MCF7 (Lane 2), A-431 (Lane 3), HaCaT (Lane 4), IMR-32 (Lane 5) and SH-SY5Y (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 (1:5000 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
- Immunofluorescent analysis of Cytokeratin 5 in methanol-fixed HeLa cells using a Cytokeratin 5 polyclonal antibody (Product # PA5-29670) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Cytokeratin 5 was performed using 70% confluent log phase A-431 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 5 Polyclonal Antibody (Product # PA5-29670) at 1:200 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
- Immunohistochemistry (Paraffin) analysis of Cytokeratin 5 was performed in paraffin-embedded human breast carcinoma tissue using Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cytokeratin 5 Polyclonal Antibody detects Cytokeratin 5 protein at cytosol and membrane on mouse intestine by immunohistochemical analysis. Sample: Paraffin-embedded mouse intestine. Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cytokeratin 5 Polyclonal Antibody detects Cytokeratin 5 protein at cytosol and membrane on mouse intestine by immunohistochemical analysis. Sample: Paraffin-embedded mouse intestine. Cytokeratin 5 Polyclonal Antibody (Product # PA5-29670) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.