Anti-CRMP1 + CRMP2 antibody [EPR7792]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human glioma tissue sections labeling CRMP2 with Purified ab129082 at 1 : 1000 dilution (0.12 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling CRMP2 with Purified ab129082 at 1 : 1000 dilution (0.12 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

Overlay histogram showing SH-SY5Y cells stained with ab129082 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129082, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

ab129082, at a dilution of 1/100, staining CRMP2 in paraffin embedded Human astrocytoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

ab129082 (purified) at 1 : 20 dilution (0.5μg) immunoprecipitating CRMP2 in U-87 MG whole cell lysate.
Lane 1 (input) : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab129082 & U-87 MG whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab129082 in U-87 MG whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-CRMP1 + CRMP2 antibody [EPR7792] (ab129082)

Predicted band size: 47 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

ab129082 staining CRMP2 in NIH/3T3 (mouse embryonic fibroblast) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

Negative control 1 : PBS only.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling CRMP2 with Purified ab129082 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Lab

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates

All lanes:

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (ab129082) at 1/50000 dilution

Lane 1:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

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: 64 kDa

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• WB

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Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

All lanes:

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (ab129082) at 1/10000 dilution

Lane 1:

PC12 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

U87 MG cell lysate at 10 µg

Lane 4:

NIH 3T3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Observed band size: 64 kDa

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• WB

Lab

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

All lanes:

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (ab129082) at 1/100000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysates

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: 64 kDa

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• WB

Lab

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

Blocking and dilution buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (ab129082) at 1/1000 dilution

Lanes 1 and 3:

Western blot - Recombinant Human CRMP1 protein (His tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-crmp1-protein-ab202599'>ab202599</a>)

Lanes 2 and 4:

Western blot - Recombinant Human CRMP2 protein (GST tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-crmp2-protein-ab114269'>ab114269</a>)

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

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Exposure time: 3s

• OI-RD Scanning

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OI-RD Scanning - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-CRMP1 + CRMP2 antibody [EPR7792] (AB129082)

CRMP1 + CRMP2 western blot using anti-CRMP1 + CRMP2 antibody [EPR7792] ab129082. Publication image and figure legend from McLean, F. H., Campbell, F. M., et al., 2019, Nutr Metab (Lond), PubMed 31168311.

ab129082 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab129082 please see the product overview.

a Representative immunoblots showing changes in DRP-2 and beta-actin protein expression in response to fatty acid free BSA, PA and OA challenge in N42 cells b Quantification of DRP-2 bands normalised to beta-actin (n = 4 plates) BSA - bovine serum albumin, DRP-2 - dihydropyrimidinase-related protein 2, PA - palmitic acid, OA - oleic acid (* p < 0.05). c-e Representative fluorescence microscopy images of N42 hypothalamic neuronal mitochondria using MitoTracker® Red CMXRos. The red colour corresponds to mitochondria. Cells were challenged with, c fatty acid free BSA, d 200 μM PA and e 200 μM OA (Bar = 10 μm. Magnification = X100). f The percentage (%) area of the cell occupied by mitochondria after challenge. BSA - bovine serum albumin, PA - palmitic acid, OA - oleic acid (*p < 0.05, **p < 0.01, ***p < 0.001)

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