702063

antibody from Invitrogen Antibodies

Targeting: NSUN5 FLJ10267, NOL1R, NSUN5A, p120(NOL1), WBSCR20, WBSCR20A, Ynl022cL

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

702063

Provider

Invitrogen Antibodies

Product name

NSUN5 Recombinant Rabbit Monoclonal Antibody (9H9L15)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

9H9L15

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

Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1), A-431 (Lane 2) and HeLa (Lane 3). The blots were probed with Anti-NSUN5 Recombinant Rabbit Monoclonal Antibody (Product # 702063, 1-2 µg/mL). A 47 kDa band corresponding to NSUN5 was observed across the cell lines 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).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of NSUN5 was achieved by transfecting HeLa cells with NSUN5 specific siRNA (Silencer® select Cat # s31270 and s224380). Western blot analysis (Fig a) was performed using Modified Whole cell extract (1 % SDS)from the NSUN5 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-NSUN5 Recombinant Rabbit Monoclonal Antibody (Product # 702063, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 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 NSUN5.