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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
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- Product number
- PA5-22027 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cdc45L Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: H1299.
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Cdc45L Polyclonal Antibody (Product # PA5-22027). Sample (30 µg of whole cell lysate). A: H1299. 7.5% SDS PAGE. Cdc45L Polyclonal Antibody (Product # PA5-22027) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Cdc45L was achieved by transfecting K562 with Cdc45L specific siRNAs (Silencer® select Product # s15830, s15829). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the Cdc45L knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cdc45L Polyclonal Antibody (Product # PA5-22027, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 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 Cdc45L.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cdc45L Polyclonal Antibody, (Product # PA5-22027) and a 60 kDa band corresponding to Cdc45L along with an uncharacterized band (*) at ~50 kDa was observed in the cell lines tested with higher expression in K-562 compared to K-562 treated with PMA which downregulates the protein expression. Modified whole cell extracts (1% SDS) (40 µg lysate) of K-562 (Lane 1), K-562 treated with PMA (16nM for 48 hours) (Lane 2), and Jurkat (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were 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
- Immunofluorescent analysis of CDC45L showing staining in the nucleus of HeLa cells. HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min and stained using a CDC45L polyclonal antibody (Product # PA5-22027) diluted at 1:500. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cdc45L Polyclonal Antibody detects CDC45L protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: CDC45L protein stained by Cdc45L Polyclonal Antibody (Product # PA5-22027) diluted at 1:500. Blue: Hoechst 33342 staining.
