Anti-Calponin 1 抗体 [EP798Y]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 1 antibody [EP798Y] (AB46794)

ICC/IF image of unpurified ab46794 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

Cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46794, 5μg/ml) overnight at +4°C. The secondary antibody (green) was DyLight^® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling Calponin 1 with purified ab46794 at a 1 : 1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used.

PBS instead of the primary antibody was used as the negative control (inset).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Immunohistochemical analysis of formalin fixed paraffin embedded human endometrium labelling calponin 1 with ab46794 at a concentration of 0.1µg/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.

ab46794 anti-Calponin 1 antibody [EP798Y] 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-Calponin 1 antibody [EP798Y] (AB46794)

Immunohistochemical staining of paraffin-embedded human smooth muscle using unpurified ab46794 at 1/100 dilution

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue sections labeling Calponin 1 with purified ab46794 at 1 : 1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used.

PBS instead of the primary antibody was used as the negative control (inset).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Paraformadehyde-fixed, 0.25% Triton X-100 permeabilized mouse thoracic aortic smooth muscle cells labeling Calponin 1 using ab46794 at 1/100 dilution in ICC/IF, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) (ab150077) at 1/400 dilution.

1.5% BSA used used as blocking agent for 30 minutes at 25°C. Incubated with primary antibody for 24 hours at 4°C.

VSMCs were seeded to 35-mm plates in a low density avoiding overlapping of cells. After fixation, VSMCs were treated with 0.25% Triton X-100 for 20 minutes.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Representative photomicrograph of mouse UT-myo cells (Left panel) and murine uterine myometrium (Right panel) stained with smooth muscle cell markers, alpha-SMA (red) and ab46794 (green) and DAPI (blue). UT-myo cells and whole-mount uterine tissue were collected from day 19 of mouse pregnancy. The placenta and embryo were removed from whole-mount tissue sections.

For full details please see paper.

Image from Herington et al PLoS One. 2015 Nov 24;10(11):e0143243. doi: 10.1371/journal.pone.0143243. eCollection 2015. Fig 1.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat lung tissue sections labeling Calponin 1 with purified ab46794 at 1 : 1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody.

PBS instead of the primary antibody was used as the negative control (inset).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Immunocytochemistry/ Immunofluorescence analysis of C2C12 (Mouse myoblasts myoblast) cells labeling Calponin 1 with purified ab46794 at 1/500 dilution. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Calponin 1 antibody [EP798Y] (ab46794) at 1/50000 dilution

Lane 1:

Human bladder lysates at 15 µg

Lane 2:

Mouse bladder lysates at 15 µg

Lane 3:

Rat bladder lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 33 kDa

false

• WB

Unknown

Western blot - Anti-Calponin 1 antibody [EP798Y] (AB46794)

All lanes:

Western blot - Anti-Calponin 1 antibody [EP798Y] (ab46794) at 1/20000 dilution

All lanes:

Human bladder lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 33 kDa

Observed band size: 34 kDa

false

• WB

AbReview25225****

Western blot - Anti-Calponin 1 antibody [EP798Y] (AB46794)

All lanes:

Western blot - Anti-Calponin 1 antibody [EP798Y] (ab46794) at 1/20000 dilution

All lanes:

Whole tissue lysate prepared from bovine lymph vessels at 10 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit polyclonal at 1/5000 dilution

Predicted band size: 33 kDa

Observed band size: 34 kDa

true

Exposure time: 1min

Image courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 showing negative staining in skeletal muscle tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 showing positive staining in normal lung vessel tissue.

• ICC/IF

AbReview18663****

Immunocytochemistry/ Immunofluorescence - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 staining Calponin 1 in porcine aortic smooth muscle cells by Immunocytochemistry/ Immunofluorescence.

The cells were paraformaldehyde fixed, permeabilized in 0.1% Triton X-100. Samples were then incubated with primary antibody at 1/50 for 1 hour at 25°C. The secondary antibody used was ab6717 Goat polyclonal to Rabbit IgG - H&L (FITC) (green) used at a 1/400 dilution.

This image is courtesy of an Abreview submitted by Jordan Carbary.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 showing positive staining in normal tonsil vessel tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 showing positive staining in normal kidney vessels tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Unpurified ab46794 showing positive staining in normal uterus tissue.

• WB

Lab

Western blot - Anti-Calponin 1 antibody [EP798Y] (AB46794)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Calponin 1 antibody [EP798Y] (ab46794) at 1/5000 dilution

All lanes:

Pig heart lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 33 kDa

false

• WB

PubMed

Western blot - Anti-Calponin 1 antibody [EP798Y] (AB46794)

ab46794 was used to stain Calponin 1 in Neonatal piglets injected with Dox (Failing LV myocardium) and PBS (Non-Failing LV myocardium). At the protein level a more than twofold increase in Calponin 1 was observed in Dox-injected animals compared to controls (PBS).

All lanes:

Western blot - Anti-Calponin 1 antibody [EP798Y] (ab46794)

Lanes 1 - 2:

Non-Failing pig LV-myocardium tissue lysate

Lanes 3 - 4:

Failing pig LV-myocardium tissue lysate injected with DOX

Predicted band size: 33 kDa

false

Torrado M et al. Targeted gene-silencing reveals the functional significance of myocardin signaling in the failing heart. PLoS One 6:e26392 (2011). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Calponin 1 antibody [EP798Y] (AB46794)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.