711274

antibody from Invitrogen Antibodies

Targeting: RHOB ARH6, ARHB, MST081, RHOH6

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711274

Provider

Invitrogen Antibodies

Product name

RhoB Recombinant Polyclonal Antibody (19HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Mouse, Monkey, Goat and Cat.

Antibody clone number

19HCLC

Concentration

0.5 mg/mL

References

Submitted references

Tumor necrosis factor-induced ArhGEF10 selectively activates RhoB contributing to human microvascular endothelial cell tight junction disruption.

Khan A, Ni W, Lopez-Giraldez F, Kluger MS, Pober JS, Pierce RW
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021 Jun;35(6):e21627

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 HeLa (Lane 1), A549 (Lane 2), COLO 205 (Lane 3), SH-SY5Y (Lane 4), Mouse Brain (Lane 5) and Rat Brain (Lane 6). The blots were probed with Anti-RhoB Recombinant Rabbit Polyclonal Antibody (Product # 711274, 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 22 kDa band corresponding to RhoB was observed across 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, UV (302 nm, 60 minutes) treated A549 cells were fixed and permeabilized for detection of endogenous RhoB using Anti-RhoB Recombinant Rabbit Polyclonal Antibody (Product # 711274, 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 RhoB 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 cytoplasmic localization of RhoB. 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.