PA5-29052

antibody from Invitrogen Antibodies

Targeting: SMC3 BAM, bamacan, CSPG6, HCAP, SMC3L1

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

PA5-29052

Provider

Invitrogen Antibodies

Product name

SMC3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, H1299, HeLa, HepG2, Molt-4, Raji, mouse liver. Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (98%), Xenopus laevis (99%), Chicken (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.95 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 using SMC3 Polyclonal Antibody (Product # PA5-29052). Sample (30 µg of whole cell lysate). A: A431. B: H1299. C: HeLa. 7.5% SDS PAGE. SMC3 Polyclonal Antibody (Product # PA5-29052) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using SMC3 Polyclonal Antibody (Product # PA5-29052). Sample (50 µg of whole cell lysate). Lane A: Mouse liver . 5% SDS PAGE. SMC3 Polyclonal Antibody (Product # PA5-29052) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of SMC3 was achieved by transfecting HeLa with SMC3 specific siRNAs (Silencer® select Products # s17427 s17426). Western blot analysis (Fig. a) was performed using whole cell extracts from the SMC3 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with SMC3 Polyclonal Antibody (Product # PA5-29052, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25ug/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SMC3. (Note: Few uncharacterized band were observed at ~80, ~50 kDa).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SMC3 Polyclonal Antibody (Product # PA5-29052) and a 160kDa band corresponding to SMC3 was observed in all the tested cell and tissue models. Modified whole cell lysate (1% SDS) (30ug lysate) of A-431 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), A549 (Lane 4), U-87 MG (Lane 5), SH-SY5Y (Lane 6), HCT 116 (Lane 7), Jurkat (Lane 8) and tissue extract (30ug lysate) of Mouse Liver (Lane 9) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by wet transfer. The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).