711866

antibody from Invitrogen Antibodies

Targeting: CDC5L CDC5, CEF1, hCDC5, PCDC5RP

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711866

Provider

Invitrogen Antibodies

Product name

Cdc5L Recombinant Polyclonal Antibody (22HCLC)

Antibody type

Polyclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Rabbit, Cat, Horse.

Antibody clone number

22HCLC

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of CDC5L was achieved by transfecting HeLa cells with CDC5L specific siRNA (Silencer® select Product # s2740+s2739). Western blot analysis (Fig a) was performed using whole cell extracts from CDC5L knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-CDC5L Recombinant Rabbit Polyclonal Antibody (Product # 711866, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to CDC5L.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HeLa (Lane 1) Hep G2 (Lane 2), U-2 OS (Lane 3), U-87 MG (Lane 4), T98G (Lane 5), HCT 116 (Lane 6) and NIH/3T3 (Lane 7). The blots were probed with Anti-CDC5L Recombinant Rabbit Polyclonal Antibody (Product # 711866, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 93 kDa band corresponding to CDC5L was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).