Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cofilin-1 with ab300185 at 1/10000 (0.104 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab300185 for 10 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Cofilin-1 with ab300185 at 1/100 (10.39 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

• sELISA

Supplier Data

Sandwich ELISA - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Sandwich ELISA of ab300183 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Cofilin-1 with ab300185 at 1/10000 (0.104 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen.
The section was incubated with ab300185 for 10 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Cofilin-1 with ab300185 at 1/100 (10.39 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in PC-12 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (ouse embryonic fibroblast) cells labelling Cofilin-1 with ab300185 at 1/100 (10.39 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Cofilin-1 with ab300185 at 1/10000 (0.104 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab300185 for 10 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IP

Lab

Immunoprecipitation - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

Cofilin-1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab300185 at 1/50 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300185 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (ab300185) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300185 in NIH/3T3 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 18 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (AB300185)

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

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Western blot - Anti-Cofilin-1 antibody [EPR24514-8] - BSA and Azide free (Detector) (ab300185) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 18 kDa

false

Exposure time: 1s