Anti-Desmin antibody [Y66] - Low endotoxin, Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (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] - Low endotoxin, Azide free (AB216616)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab32362).

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] - Low endotoxin, Azide free (AB216616)

This data was developed using the same antibody clone in a different buffer formulation (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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (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] - Low endotoxin, Azide free (AB216616)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab32362).

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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab32362).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab32362).

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

Clone Y66 (ab216616) has been successfully conjugated by Abcam. This image was generated using Anti-Desmin antibody [Y66] - Cytoskeleton Marker (Alexa Fluor® 647). Please refer to ab195177 for protocol details.

ab195177 staining Desmin in A673 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes 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 ab195177 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in A673 cells fixed with 100% methanol (5 min)

• ICC/IF

AbReview38459****

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

This image is courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab32362).

• IHC-P

AbReview25190****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

Unpurified ab32362 staining Desmin in nude rat esophagheal tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6. Samples were then blocked with 1% BSA for 20 minutes at 25°C and then incubated with unpurified ab32362 at a 1/400 dilution for 16 hours at 25°C. The secondary used was an undiluted goat anti-rabbit HRP conjugated polyclonal.Striated muscle cells of muscular propria are strongly positive for desmin. Smooth muscle cells and vascular smooth muscle cells in submucosal layer are also positive for desmin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

This image is courtesy of an anonymous Abreview.

• ICC/IF

AbReview38457****

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

This image is courtesy of an anonymous Abreview.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

• WB

Lab

Western blot - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

All lanes:

Western blot - Anti-Desmin antibody [Y66] - Cytoskeleton Marker (<a href='/en-us/products/primary-antibodies/desmin-antibody-y66-cytoskeleton-marker-ab32362'>ab32362</a>) 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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

Clone Y66 (ab216616) has been successfully conjugated by Abcam. This image was generated using Anti-Desmin antibody [Y66] - Cytoskeleton Marker (Alexa Fluor® 488). Please refer to ab185033 for protocol details.

ab185033 staining Desmin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab185033 at 1/50 dilution overnight at +4°C (shown in green). AlexaFluor^®350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32362).

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

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Desmin antibody [Y66] - Low endotoxin, Azide free (AB216616)

Clone Y66 (ab216616) has been successfully conjugated by Abcam. This image was generated using Anti-Desmin antibody [Y66] - Cytoskeleton Marker (PE). Please refer to ab224935 for protocol details.

Overlay histogram showing SV40LT-SMC cells stained with ab224935 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab224935, 1/1000 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

This antibody gave a positive signal in SV40LT-SMC cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.