Anti-CKAP4 抗体 [EPR25713-62] (ab302539)

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

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

The band beneath the target band (63 kDa) is likely to be degraded target fragments.

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

Lysates were freshly made and used immediately. 

Blocking and dilution 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.

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling CKAP4 with ab302539 at 1/2000 dilution (0.255 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling CKAP4 with ab302539 at 1/2000 dilution (0.255 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CKAP4 with ab302539 at 1/2000 dilution (0.255 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling CKAP4 with ab302539 at 1/2000 dilution (0.255 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling CKAP4 with ab302539 at 1/2000 dilution (0.255 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling CKAP4 with AB302539 at 1/50 dilution (10.18 µg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preabsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in HeLa cell line. Ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary diluent was used instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preabsorbed at 1/1000 dilution (2 µg/mL).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling CKAP4 with AB302539 at 1/50 (10.18 µg/mL) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preabsorbed antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line. Ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^®594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody control: Primary diluent was used instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preabsorbed at 1/1000 dilution (2 µg/mL).

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling CKAP4 with AB302539 at 1/500 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled 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.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling CKAP4 with AB302539 at 1/500 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled 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.

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.

Exposure time: 3 seconds. 

Observed MW (kDa): 63. 

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.

Exposure time: 3 seconds

 Observed MW (kDa): 63.