MA1-25492

antibody from Invitrogen Antibodies

Targeting: YWHAZ 14-3-3-zeta, KCIP-1, YWHAD

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-25492

Provider

Invitrogen Antibodies

Product name

14-3-3 beta/zeta Monoclonal Antibody (22-IID8B)

Antibody type

Monoclonal

Antigen

Other

Description

Recommended positive controls: HeLa cell lysate.

Antibody clone number

22-IID8B

Concentration

1 mg/mL

References

Submitted references

A combined proteome and ultrastructural localization analysis of 14-3-3 proteins in transformed human amnion (AMA) cells: definition of a framework to study isoform-specific differences.

Moreira JM, Shen T, Ohlsson G, Gromov P, Gromova I, Celis JE
Molecular & cellular proteomics : MCP 2008 Jul;7(7):1225-40

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of human epidermoid carcinoma A431 cell lysate, probed with 14-3-3 beta/zeta Monoclonal Antibody (22-IID8B) (Product # MA1-25492).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of 14-3-3 protein beta/zeta was achieved by transfecting MCF7 with 14-3-3 protein beta/zeta specific siRNAs (Silencer® select Product # s14961, s14962, s14972). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the 14-3-3 protein beta/zeta knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with 14-3-3 beta/zeta Monoclonal Antibody (22-IID8B) (Product # MA1-25492, 1:1000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 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 14-3-3 protein beta/zeta.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-14-3-3 beta/zeta Monoclonal Antibody (22-IID8B) (Product # MA1-25492) and a 28 kDa band corresponding to 14-3-3 protein beta/zeta was observed across the cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), PC-3 (Lane 3), A549 (Lane 4), Mouse Skeletal Muscle (Lane 5), Rat Skeletal Muscle (Lane 6), Mouse Brain (Lane 7), Rat Brain (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).