702005

antibody from Invitrogen Antibodies

Targeting: MECOM EVI1, KMT8E, MDS1, MDS1-EVI1, PRDM3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702005

Provider

Invitrogen Antibodies

Product name

EVI1 Recombinant Rabbit Monoclonal Antibody (11H41L13)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

11H41L13

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), HEK-293 (Lane 2), A-431 (Lane 3), PC-3 (Lane 4), Caco-2 (Lane 5), HEL 92.1.7 (Lane 6) and U-2 OS (Lane 7). The blots were probed with Anti-EVI1/ MECOM Recombinant Rabbit Monoclonal Antibody (Product # 702005, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 120 kDa band corresponding to EVI1/ MECOM was observed across the cell lines tested. 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, Retinoic acid (10 uM, 48 h) treated NTERA-2 cells were fixed and permeabilized for detection of EVI1/ MECOM using Anti-EVI1/ MECOM Recombinant Rabbit Monoclonal Antibody (Product # 702005, 2 µg/mL) 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 EVI1/ MECOM protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of EVI1/ MECOM. Panel e) shows untreated cells with no signal. Panel f) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.