Anti-TAGLN/Transgelin 抗体

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TAGLN/Transgelin antibody (AB14106)

ab14106 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed in 100% methanol for 5 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14106 at 1 μg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor^® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature.

DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1 hour at room temperature.

• ICC/IF

AbReview42536****

Immunocytochemistry/ Immunofluorescence - Anti-TAGLN/Transgelin antibody (AB14106)

ab14106 staining SM22 alpha in mouse muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence).

Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/1000.

This image is courtesy of an anonymous Abreview.

• WB

Ap

Western blot - Anti-TAGLN/Transgelin antibody (AB14106)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14106 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/mL

Lane 1:

Human colon tissue lysate - total protein (ab30051) at 10 µg

Lane 2:

Mouse colon tissue lysate at 10 µg

Lane 3:

Rat colon tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Predicted band size: 23 kDa

Observed band size: 115 kDa,20 kDa,23 kDa,36 kDa

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Exposure time: 5s

• WB

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Western blot - Anti-TAGLN/Transgelin antibody (AB14106)

The band was completed abolished by peptide blocking with ab16067 (immunizing peptide) - not shown.

All lanes:

Western blot - Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/mL

Lane 1:

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

Lane 2:

HeLa nuclear extract at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

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• WB

Unknown

Western blot - Anti-TAGLN/Transgelin antibody (AB14106)

All lanes:

Western blot - Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/mL

Lane 1:

Western blot - Recombinant Human TAGLN/Transgelin protein (His tag N-Terminus) (<a href='/products/proteins-peptides/recombinant-human-tagln-transgelin-protein-ab101469'>ab101469</a>) at 0.01 µg

Lane 1:

Western blot - Recombinant Human SPG21/Maspardin protein (His tag N-Terminus) (<a href='/products/proteins-peptides/recombinant-human-spg21-maspardin-protein-ab86438'>ab86438</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 23 kDa,35 kDa

true

Exposure time: 30s

• WB

AbReview16283****

Western blot - Anti-TAGLN/Transgelin antibody (AB14106)

All lanes:

Western blot - Anti-TAGLN/Transgelin antibody (ab14106) at 1/1000 dilution

All lanes:

Lysates prepared from human primary smooth muscle cells at 30 µg

Secondary

All lanes:

HRP-conjugated bovine polyclonal to rabbit IgG at 1/2000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

true

Exposure time: 2min

This image is a courtesy of Anonymous Abreview

• WB

CiteAb

Western blot - Anti-TAGLN/Transgelin antibody (AB14106)

Western Blotting using Anti-TAGLN/Transgelin antibody, ab14106. Publication image from Boehm, M. et al., 2016, Nat Commun, 27340017. Legend direct from paper.

Hypoxia increases EndMT and is additive to the effect of TGF-β.(a) Representative pimonidazole staining demonstrating the extent of hypoxia in aortic sections from tamoxifen-induced end.SclCreERT;R26RstopYfp;ApoE−/− mice fed 8 versus 18 weeks HFD. Staining in red is indicative of hypoxia with pO2 <10 mm Hg. Specific staining was performed using pimonidazole (seen in red) and DAPI (blue). L=lumen; scale bars, 100 µm. Staining was performed on at least three spatially separated thoracic aortic sections per mouse (n=5 mice per group). Analysis by one-way ANOVA; overall P<0.0001. (b) Immunostaining of HCAECs exposed to hypoxia and/or TGF-β for 5 days in vitro. Scale bars, 100 µm. (c–f) Relative RNA expression of mesenchymal genes FAP, SM22α, Calponin and Versican (respectively) in HCAECs was increased by hypoxia as assessed by qRT-PCR. (g–j) Relative RNA expression of endothelial genes CD31, Ve-Cadherin, eNOS and KDR assessed by qRT-PCR showed mixed effects in response to hypoxia. (k–l) Relative RNA expression of SNAI2 and SNAI1 in HCAECs was increased by hypoxia. (m) SM22α protein expression, assessed by western blot in HCAECs, was increased by hypoxia as indicated. SM22α ladder (L) represents 25 kDa (upper), 20 kDa (middle) and 15 kDa (lower). SM22α expected molecular weight/size is 23 kDa. GAPDH ladder represents 37 kDa. GAPDH expected molecular weight/size is 37 kDa. (n) CD31 protein expression, assessed by western blot in HCAECs. CD31 ladder (L) represents 150 kDa. CD31 expected molecular weight/size is 130 kDa. GAPDH ladder represents 37 kDa. GAPDH expected molecular weight/size is 37 kDa. As our intent was to evaluate the specific effect of hypoxia, and not interactions with TGF-β (which is provided as a reference), t testing was performed in this case. n=3 in all experiments. All experiments replicated >3 times. NS, not significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

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