Anti-CKAP4 antibody [EPR25713-62]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CKAP4 with ab302539 at 1/50 (10.18 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling CKAP4 with ab302539 at 1/2000 (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human lung cancer (PMID : 29751934). The section was incubated with ab302539 for 30 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CKAP4 with ab302539 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CKAP4 with ab302539 at 1/2000 (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human colon. The section was incubated with ab302539 for 30 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunohistochemical analysis of paraffin-embedded Human cerebral corte tissue labeling CKAP4 with ab302539 at 1/2000 (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human cerebral cortex. The section was incubated with ab302539 for 30 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

CKAP4 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab302539 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302539 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg (Inset)
Lane 2 : ab302539 IP in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab302539 in HeLa whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/30 dilution

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

Predicted band size: 66 kDa

Observed band size: 63 kDa

false

Exposure time: 3s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CKAP4 with ab302539 at 1/50 (10.18 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral corte tissue labeling CKAP4 with ab302539 at 1/2000 (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebral cortex. The section was incubated with ab302539 for 30 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling CKAP4 with ab302539 at 1/2000 (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat cerebral cortex. The section was incubated with ab302539 for 30 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CKAP4 with ab302539 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

CKAP4 was immunoprecipitated from 0.35 mg (mouse embryonic fibroblast), whole cell lysate with ab302539 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302539 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 μg (Inset)
Lane 2 : ab302539 IP in NIH/3T3 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab302539 in NIH/3T3 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/30 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast), 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

Predicted band size: 66 kDa

Observed band size: 63 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

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

All lanes:

Western blot - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting cytoskeleton-associated protein 4, whole cell lysate 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 1/100000 dilution

Observed band size: 63 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used immediately.

All lanes:

Western blot - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

A549 (human lu carcinoma epithelial cell), whole cell lysate 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 1/100000 dilution

Observed band size: 60 kDa,63 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : human skeletal muscle (HPA database). The band beneath the target band (63 kDa) is likely to be degraded target fragments.

All lanes:

Western blot - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/1000 dilution

Lane 1:

Human colon tissue lysate 20 µg

Lane 2:

Human colon cancer tissue lysate 20 µg

Lane 3:

Human lu cancer tissue lysate 20 µg

Lane 4:

Human skeletal muscle lysate 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/5000 dilution

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-CKAP4 antibody [EPR25713-62] (AB302539)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The band beneath the target band (63 kDa) is likely to be degraded target fragments.

All lanes:

Western blot - Anti-CKAP4 antibody [EPR25713-62] (ab302539) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate 20 µg

Lane 2:

3T3-L1 (mouse embryonic fibroblast), whole cell lysate 20 µg

Lane 3:

Mouse pancreas cancer tissue lysate 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 1/100000 dilution

Observed band size: 63 kDa

false

Exposure time: 10s