Anti-CRMP1 antibody [EP14521] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CRMP1 with ab199722 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. No staining on Human liver tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling CRMP1 with ab199722 at 1/20 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) and Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling CRMP1 with ab199722 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm and nuclear staining on U-87 MG cell line is observed. Negative expression in Jurkat cells. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows : -
-ve control 1 : ab199722 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling CRMP1 with ab199722 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nucleus staining on Human cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

CRMP1 was immunoprecipitated from 1mg of Human fetal brain whole cell lysate with ab199722 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199722 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

Lane 1 : Human fetal brain whole cell lysate 10ug (Input). Lane 2 : ab199722 IP in Human fetal brain whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199722 in Human fetal brain whole cell lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-CRMP1 antibody [EP14521] (<a href='/en-us/products/primary-antibodies/crmp1-antibody-ep14521-ab199722'>ab199722</a>)

Predicted band size: 62 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling CRMP1 with ab199722 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on rat cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling CRMP1 with ab199722 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Based on the sequence analysis, ab199722 recognizes two isoforms with the predicted MWs of 62 kDa and 74 kDa, respectively.

All lanes:

Western blot - Anti-CRMP1 antibody [EP14521] (<a href='/en-us/products/primary-antibodies/crmp1-antibody-ep14521-ab199722'>ab199722</a>) at 1/5000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

SH-SY5Y (Human neuroblastoma from bone marrow cells) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa,62 kDa

Observed band size: 52 kDa,64 kDa,80 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

Blocking buffer : 5% NFDM/TBST.

Dilution buffer : 5% NFDM /TBST or 1% BSA/TBST.

Based on the sequence analysis, CRMP1 does not have similar isoforms within mouse and rat, therefore only a single band at approximately 62kDa is seen in mouse and rat lysates within western blot.

All lanes:

Western blot - Anti-CRMP1 antibody [EP14521] (<a href='/en-us/products/primary-antibodies/crmp1-antibody-ep14521-ab199722'>ab199722</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

C6 (Rat glial tumor cells) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 62 kDa

Observed band size: 64 kDa

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

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

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CRMP1 antibody [EP14521] (<a href='/en-us/products/primary-antibodies/crmp1-antibody-ep14521-ab199722'>ab199722</a>) at 1/1000 dilution

All lanes:

N-His tagged Recombinant Human CRMP1 protein (aa 1 to 572) at 0.01 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

Predicted band size: 62 kDa

Observed band size: 65 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-CRMP1 antibody [EP14521] - BSA and Azide free (AB251288)

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

This data was developed using ab199722, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CRMP1 antibody [EP14521] (<a href='/en-us/products/primary-antibodies/crmp1-antibody-ep14521-ab199722'>ab199722</a>) at 1/1000 dilution

All lanes:

N-GST tagged Recombinant Human CRMP2 protein (aa1-572) at 0.01 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

Predicted band size: 62 kDa

Observed band size: 89 kDa

false

Exposure time: 3s