Anti-Desmin antibody [Y66] - Cytoskeleton Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunofluorescence staining of Desmin using ab32362 in ab277612, which were differentiated for 10 days post induction.

The cells were fixed with 100% MeOH (5 min) and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32362 at 0.02 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.

The antibody ab32362 gave comparable results using 4% formaldehyde fixation (10 min).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human urinary bladder tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human tonsil tissue. Unpurified ab32362 shows negative staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemical analysis of formalin fixed paraffin embedded human heart labelling Desmin with ab32362 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

Anti-Desmin antibody [Y66] ab32362 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling Desmin with purified ab32362 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control : primary antibody (1/50) and secondary antibody ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human skeletal muscle tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human uterus tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

AbReview38459****

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Unpurified ab32362 staining Desmin (green) in Human skeletal muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methacarn and blocked with 10% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/150) for 12 hours. An Alexa Fluor^® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Blue - DAPI-nuclei. Red - WGA. 40X objective.

This image is courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labelling Desmin with purified ab32362 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human brain tissue. Unpurified ab32362 shows negative staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunocytochemistry/Immunofluorescence analysis of C2C12 (Mouse myoblasts myoblast) cells labeling Desmin with ab32362 at 1/500. Cells were fixed with 100% Methanol. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 was used as counterstain antibody.

Confocal image showing cytoplasmic staining on C2C12 cell line.

• ICC/IF

AbReview38457****

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Unpurified ab32362 staining Desmin (green) in Mouse aorta smooth muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formalin and blocked with 10% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/150) for 1 hour at 22°C. An Alexa Fluor^® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Blue - nuclei.

This image is courtesy of an anonymous Abreview.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Intracellular Flow Cytometry analysis of C2C12 cells labelling Desmin with purified ab32362 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• WB

AbReview38458****

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362) at 1/500 dilution

All lanes:

Human skeletal muscle whole tissue lysate at 20 µg

Secondary

All lanes:

IRDye® 680-conjugated anti-rabbit at 1/5000 dilution

Predicted band size: 54 kDa

Observed band size: 53 kDa

true

Exposure time: 50s

This image is courtesy of an anonymous Abreview

• WB

Lab

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Blocking and diluting buffer concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 26041514, 32322014, and 28955862).

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 54 kDa

Observed band size: 53 kDa,35-50 kDa

false

Exposure time: 3s

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Immunofluorescent analysis of Human mitochondria injected rabbit hearts sections stained for Desmin (Green) using ab32362. MTCO2, the human-specific mitochondrial marker was stained in red, and the nuclei was stained using the DNA stain DAPI (blue).

Image from Cowan DB et al. Intracoronary Delivery of Mitochondria to the Ischemic Heart for Cardioprotection. PLoS One 11:e0160889 (2016).

• WB

Collaborator

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362) at 1/100000 dilution

Lane 1:

Guinea pig heart tissue lysate at 10 µg

Lane 2:

Guinea pig muscle tissue lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 54 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362) at 1/500000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 54 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362) at 1/500000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

Human fetal heart tissue lysate at 20 µg

Lane 3:

Human fetal muscle tissue lysate at 20 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 54 kDa

Observed band size: 53 kDa

false

• WB

CiteAb

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (AB32362)

Desmin western blot using anti-Desmin antibody [Y66] - Cytoskeleton Marker ab32362. Publication image and figure legend from Xu, M., Zhang, F., et al., 2016, PLoS One, PubMed 27907201.

ab32362 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab32362 please see the product overview.

TWEAK significantly increased the expression of α-SMA, vimentin, desmin in LX-2 cells and changed the cell morphology.The mRNA (A) and protein (B) of α-SMA, vimentin, desmin in LX-2 cells treated with 40 ng/ml and 100 ng/ml TWEAK for 24 h were analyzed by qRT-PCR and western blot. β-actin served as an internal control. Data are presented as mean ± SD of three independent experiments. (C) Phalloidin stained actin cytoskeleton (green) in LX-2 cells. The nuclear was stained by DAPI (blue). The data shown here are from three independent experiments with similar results. Scale bars, 50um. *p<0.05,**p<0.005,***p<0.0005,****p<0.0001.

false