703394

antibody from Invitrogen Antibodies

Targeting: ACTR2 ARP2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

703394

Provider

Invitrogen Antibodies

Product name

Arp2 Recombinant Rabbit Monoclonal Antibody (5H2L7)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Antibody clone number

5H2L7

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 SH-SY5Y (Lane 1), PANC-1 (Lane 2), HeLa (Lane 3), U-87 MG (Lane 4), MDA-MB-231 (Lane 5), 3T3-L1 (Lane 6), Jurkat (Lane 7), HEK-293 (Lane 8) and tissue extract of Mouse Brain (Lane 9). The blot was probed with Anti-Arp2 Recombinant Rabbit Monoclonal Antibody (Product # 703394, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). A ~44 kDa band corresponding to Arp2 was observed across the cell lines and tissue tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of ARP2 was achieved by transfecting SH-SY5Y cells with specific siRNA (Silencer® select Product # s223082, s19644). Immunofluorescence analysis was performed on SH-SY5Y cells (untransfected, panel a-d), transfected with ARP2 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labeled with Anti-ARP2 Recombinant Rabbit Monoclonal Antibody (Product # 703394, 1:100 dilution), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Significant reduction of the signal was observed upon siRNA mediated knockdown (panel i-l) confirming the specificity of the antibody to ARP2 (green). The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, SH-SY5Y cells were fixed and permeabilized for detection of endogenous ARP2 using Anti-ARP2 Recombinant Rabbit Monoclonal Antibody (Product # 703394, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of ARP2 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of ARP2. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.