Anti-CRMP2 antibody [EPR28480-66]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SK-BR-3 (human breast adenocarcinoma epithelial cell, Left) / SH-SY5Y (human neuroblastoma epithelial cell, Right) cells labelling CRMP2 with ab315285 at 1/5000 dilution (0.01 ug, 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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low or negative expression : SK-BR-3 (PMID : 32778769).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling CRMP2 with ab315285 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on human cerebrum. The section was incubated with ab315285 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling CRMP2 with ab315285 at 1/1000 (0.503 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in SH-SY5Y cell line.Low or negative expression : SK-BR-3 (PMID : 32778769). 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 ug/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 (2 ug/ml) dilution.

• IP

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Immunoprecipitation - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

CRMP2 was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate with ab315285 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315285 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

Lane 2 : ab315285 IP in SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315285 in SH-SY5Y whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CRMP2 antibody [EPR28480-66] (ab315285) at 1/30 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell) 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

false

Exposure time: 3s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling CRMP2 with ab315285 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat kidney. The section was incubated with ab315285 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).

Heat mediated antigen retrieval was performed 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-CRMP2 antibody [EPR28480-66] (AB315285)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CRMP2 with ab315285 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on mouse cerebrum. The section was incubated with ab315285 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling CRMP2 with ab315285 at 1/1000 (0.503 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4 ug/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 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CRMP2 with ab315285 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining on rat cerebrum. The section was incubated with ab315285 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).

Heat mediated antigen retrieval was performed 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-CRMP2 antibody [EPR28480-66] (AB315285)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling CRMP2 with ab315285 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney. The section was incubated with ab315285 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).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling CRMP2 with ab315285 at 1/1000 (0.503 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4 ug/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 (2 ug/ml) dilution.

• WB

Supplier Data

Western blot - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

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

Low or negative expression : K-562, SK-BR-3 (PMID : 32778769).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CRMP2 antibody [EPR28480-66] (ab315285) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

C6 (rat glial tumor glial 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 1/100000 dilution

Observed band size: 62 kDa,36 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

Western blot : Anti-DPYSL2 antibody [EPR28480-66] (ab315285) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab315285 was shown to bind specifically to DPYSL2. A band was observed at 63 kDa in wild-type A549 cell lysates with no signal observed at this size in DPYSL2 knockout cell line. To generate this image, wild-type and DPYSL2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CRMP2 antibody [EPR28480-66] (ab315285) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

DPYSL2 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A549 Nuclear Fraction cell lysate at 20 µg

Lane 5:

Recombinant Human CRMP2 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-crmp2-protein-ab114269'>ab114269</a>) cell lysate at 0.2 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 63 kDa

false

• WB

Supplier Data

Western blot - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CRMP2 antibody [EPR28480-66] (ab315285) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human hypothalamus tissue lysate at 20 µg

Lane 3:

Mouse hippocampus tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Rat hippocampus tissue lysate at 20 µg

Lane 6:

Rat brain tissue 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 1/100000 dilution

Observed band size: 62 kDa,36 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-CRMP2 antibody [EPR28480-66] (AB315285)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-CRMP2 antibody [EPR28480-66] (ab315285) at 1/1000 dilution

Lane 1:

His-tagged human DPYSL1 protein at 10 ng

Lane 2:

His-tagged human DPYSL2 protein at 10 ng

Lane 3:

His-tagged human DPYSL3 protein at 10 ng

Lane 4:

His-tagged human DPYSL4 protein at 10 ng

Lane 5:

His-tagged human DPYSL5 protein at 10 ng

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: 62 kDa

false

Exposure time: 37s