MA1-26058

antibody from Invitrogen Antibodies

Targeting: TGFB1 CED, DPD1, TGFB, TGFbeta

Western blot (Recommended by provider)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-26058

Provider

Invitrogen Antibodies

Product name

TGF beta-1 Monoclonal Antibody (2C5)

Antibody type

Monoclonal

Antigen

Other

Description

Western blotting demonstrated that this antibody reacts with the dimeric (25kDa) forms of TGF beta under both non-reducing and reducing conditions respectively.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

2C5

Vial size

250 µg

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of TGF beta1 (Transforming growth factor beta-1 proprotein) was performed using THP-1 and THP-1 differentiated and polarized to M2 macrophage cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TGF beta-1 Monoclonal Antibody (2C5) (Product # MA1-26058) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500 dilution), 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 increased cytoplasmic localization of TGF beta1 in M2 macrophages compared to THP-1 control cells (Panel e). Panel f represents control M2 macrophage cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of TGF beta1 was achieved by transfecting THP-1 cells with TGF beta1 specific siRNA (Silencer® select Product # s14056, s14054). Immunofluorescence analysis was performed on untransfected THP1 cells (panel a,b), untransfected M2 macrophage (panel c, d), non-specific scrambled siRNA transfected THP-1 (panels e,f), non-specific scrambled siRNA transfected M2 macrophage (panels g,h), THP-1 transfected with TGF beta1 specific siRNA (panel i,j), and THP-1 transfected with TGF beta1 specific siRNA and differentiated to M2 macrophage (panels k,l) (Green). Cells were fixed, permeabilized, and labelled with TGF beta-1 Monoclonal Antibody (2C5) (Product # MA1-26058, 1:100) followed by Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (Blue) staining. Reduction in specific signal was observed upon siRNA mediated knockdown (panel k,l) confirming specificity of the antibody to TGF beta1. The Images were captured at 60X magnification.