Anti-alpha smooth muscle Actin antibody

• IHC-P

AbReview79885****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of 10% formalin-fixed Human transitional cell carcinoma of the kidney. Stained with ab5694 at 1/150 dilution. Secondary antibody used was goat anti-rabbit Alexa-568® at 1/500 dilution. Blocking was done with 5% serum for 1 hour at 25°C. The sample was incubated with the primary antibody for 1hour at 25°C with 5% normal goat serum. Antigen retrieval method was heat mediated with EDTA pH 8.

This image is courtesy of an Abreview submitted by Cameron Johnstone.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling alpha smooth muscle Actin with ab5694 at a dilution of 1/1000. Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. Sections were incubated with the primary antibody for 1 hour followed by incubation with a biotinylated secondary antibody (ab6720) for 30 minutes then HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin.

Magnification : left - 10X, right - 40X.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

Representative histology of aortic valve leaflets from aged mice demonstrates changes in pRb cKO AoV

A) Masson's trichrome showing reduced collagen staining (blue) in leaflet from pRb cKO mouse with aortic regurgitation (AR). B) Movat pentachrome showing more diffuse collage staining (yellow) in fibrosa, but normal proteoglycan staining (blue) in the spongiosa layer of the leaflet from pRb cKO with AR. C) Immunohistochemistry for α-SMA, demonstrating presence of activated myofibroblasts throughout leaflets of pRb cKO mouse with and without AR. Scale bar is 50μm.

Alpha smooth muscle Actin is detected witn ab5694 at 1/1000 dilution.

(From Figure 2 of Freytsis et al)

Freytsis, M. et al PLoS One. 2018 Jan 5;13(1):e0190623. doi: 10.1371/journal.pone.0190623. eCollection 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1 : 1600 to 1 : 3200, incubation overnight at 4^oC, counterstained with Hematoxylin.

This image was kindly supplied as part of the review by JQ Zhang.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

Passive CLARITY technique (PACT)-sorbitol refractive index matching solution (sRIMS) clearing and 3D imaging of virgin and lactating mouse mammary tissue. a PACT-sRIMS tissue clearing and immunostaining protocol and timeline. Three-dimensional confocal imaging of PACT-sRIMS-cleared virgin (b) and lactating (c) mammary glands immunostained with basal cell markers (K5 and smooth muscle actin (stained with ab5694)) and luminal cell markers (K8 and E-cadherin (E-CAD)). Main image shows the maximum intensity projection of the entire image sequence, with thin optical slices (1 μm) and their depth (z value) relative to the first image in the image sequence.

From a paper comparing imaging of intact virgin and lactating mammary glands using 3D imaging of solvent-cleared organs, see deep brain (seeDB), clear unobstructed brain imaging cocktails (CUBIC) and passive clarity technique.

Image from Lloyd-Lewis B. et al., Breast Cancer Res. 2016 18: 127. Fig 2.; doi: 10.1186/s13058-016-0754-9. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694)

Pancreatic vessel imaging in the intact adult mouse pancreas. In adult mouse tissues (12 weeks old), imaging was performed after CLARITY. Three-dimensional projection clarified mouse pancreas with capillary immunostained for α-smooth muscle actin (green). Scale bar, 200 μm.

From a study, that aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs.

Image from Lee H. et al., BMC Dev Biol. 2014 Dec 21;14:48. Fig 2Bdoi: 10.1186/s12861-014-0048-3 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• WB

Unknown

Western blot - Anti-alpha smooth muscle Actin antibody (AB5694)

Please note that ab5694 does not appear to be specific to smooth muscle.

All lanes:

Western blot - Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/mL

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear cell lysate at 20 µg

Lane 2:

HeLa whole cell lysate at 20 µg

Lane 3:

A431 (Human epidermoid carcinoma cell line) cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

Lane 5:

HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg

Secondary

All lanes:

Alexa Fluor anti-rabbit at 1/5000 dilution

Predicted band size: 42 kDa

Observed band size: 30 kDa,35 kDa,37 kDa,42 kDa,50 kDa,75 kDa

false

• WB

Supplier Data

Western blot - Anti-alpha smooth muscle Actin antibody (AB5694)

Incubated with the primary antibody at 4°C overnight.

Incubated with the secondary antibody at room temperature for 1 hour.

Lanes 1 - 3:

Western blot - Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/mL

Lanes 4 - 5:

Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate - overexpressing alpha-Actin at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate at 20 µg

Lanes 3 and 5:

Mouse heart tissue homogenate at 20 µg

Lane 4:

NIH/3T3 cell lysate at 20 µg

Secondary

All lanes:

Fluor 750-conjugated goat anti-rabbit IgG (H+L) at 1/12500 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false