711470

antibody from Invitrogen Antibodies

Targeting: HDAC3 HD3, KDAC3, RPD3, RPD3-2

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711470

Provider

Invitrogen Antibodies

Product name

HDAC3 Recombinant Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human, Mouse, Rat

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 HDAC3 was achieved by transfecting HeLa cells with HDAC3 specific validated siRNA (Silencer® select Product # s16877). Western blot analysis was performed using whole cell lysates from the HDAC3 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-HDAC3 Recombinant Rabbit Polyclonal Antibody (Product # 711470, 1-2 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Reduction of signal upon siRNA mediated knock down confirms that antibody is specific to HDAC3.

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), C2C12 (Lane 2), COS-7 (Lane 3), Hep G2 (Lane 4), A-431 (Lane 5), MCF7 (Lane 6) and Tissue extracts of Mouse Brain (Lane 7), Rat Brain (Lane 8) and Mouse Liver (Lane 9). The blots were probed with Anti-HDAC3 Recombinant Rabbit Polyclonal Antibody (Product # 711470, 1-2 µg/mL). A 48 kDa band corresponding to HDAC3 was observed across the cell lines and tissues tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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).