Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
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- Product number
- PA5-20764 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cdc42 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control is human brain tissue lysate. PA5-20764 can be used with blocking peptide PEP-0878.
- Reactivity
- Human, Mouse, Rat
- Host
- Chicken/Avian
- Isotype
- IgY
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of human brain tissue lysate using a CDC42 polyclonal antibody (Product # PA5-20764) at (A) 0.5 and (B) 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of CDC42 in human brain tissue lysate with Cdc42 Polyclonal Antibody (Product # PA5-20764) at (A) 0.5 and (B) 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of Cdc42 was achieved by transfecting A-431 with Cdc42 specific siRNAs (Silencer® select Product # s2765, s2766). Western blot analysis (Fig. a) was performed using whole cell extracts from the Cdc42 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Cdc42 Polyclonal Antibody (Product # PA5-20764, 0.5µg/mL) and Goat anti-Chicken IgY (H+L) Secondary Antibody, HRP (Product # A16054, 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 Cdc42.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cdc42 Polyclonal Antibody (Product # PA5-20764) and 21 kDa band corresponding to Cdc42 was observed across cell lines and tissues tested along with certain uncharacterized bands at ~ 80 kDa. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), U-937 (Lane 2), A-431 (Lane 3), K-562 (Lane 4), HeLa (Lane 5), Hep G2 (Lane 6), tissue extracts of Mouse Brain (Lane 7) and Rat Brain (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5µg/mL) and detected by chemiluminescence Goat anti-Chicken IgY (H+L) Secondary Antibody, HRP (Product # A16054, 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 cdc42 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 2% BSA for 1 hour at room temperature. The cells were labeled with Cdc42 Polyclonal Antibody (Product # PA5-20764) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Chicken IgY (H+L) Secondary Antibody, Alexa Fluor 488 (Product # A-11039) 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 cytoskeletal and cytoplasmic localization. Panel e represents cells with no primary antibody to assess background. The images were captured at 60X magnification.