720394

antibody from Invitrogen Antibodies

Targeting: DTX3L BBAP, RNF143

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

720394

Provider

Invitrogen Antibodies

Product name

DTX3L Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of DTX3L was achieved by transfecting PC-3 cells with DTX3L specific siRNA (Silencer® select Product # s45594 + s45595). Western blot analysis (Fig a) was performed using Whole cell extracts from the DTX3L knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-DTX3L Rabbit Polyclonal Antibody (Product # 720394, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 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 DTX3L.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), DU 145 (Lane 2), LNCaP (Lane 3), HeLa (Lane 4), MCF-7 (Lane 5), A-431 (Lane 6), HT-29 (Lane 7) and K-562 (Lane 8). The blots were probed with Anti-DTX3L Rabbit Polyclonal Antibody (Product # 720394, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 dilution). A 83 kDa band corresponding to DTX3L was observed across the cell lines tested. Along with the desired band, non-specific band at 50 kDa was observed in 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).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), DU 145 (Lane 2), LNCaP (Lane 3), HeLa (Lane 4), MCF-7 (Lane 5), A-431 (Lane 6), HT-29 (Lane 7) and K-562 (Lane 8). The blots were probed with Anti-DTX3L Rabbit Polyclonal Antibody (Product # 720394, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 dilution). A 83 kDa band corresponding to DTX3L was observed across the cell lines tested. Along with the desired band, non-specific band at 50 kDa was observed in 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).