MA1-25057

antibody from Invitrogen Antibodies

Targeting: TUBB1 dJ543J19.4

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-25057

Provider

Invitrogen Antibodies

Product name

beta Tubulin 1,2 Monoclonal Antibody (JDR.3B8)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: chicken fibroblasts, chicken brain.

Antibody clone number

JDR.3B8

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-beta Tubulin 1,2 Monoclonal Antibody (JDR.3B8) (Product # MA1-25057) and a 50 kDa band corresponding to Tubulin beta-2A chain was observed across all the tested cell lines and tissues, except HeLa, HL-60, THP-1, Mouse Ovary and Rat Ovary. Whole cell extracts (30 µg lysate) of NTERA-2 cl.D1 (Lane 1), Caco-2 (Lane 2), BJ (Lane 3), HeLa (Lane 4), HL-60 (Lane 5), THP-1 (Lane 6), Mouse Brain (Lane 7), Mouse Ovary (Lane 8), Rat Brain (Lane 9), Rat Ovary (Lane 10) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Tubulin beta-2A chain was performed using 70% confluent log phase NTERA-2 cl.D1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with beta Tubulin 1,2 Monoclonal Antibody (JDR.3B8) (Product # MA1-25057) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoskeletal localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.