- PA5-117919 - Invitrogen Antibodies DPYSL2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot of CRMP2 in Lane A: Jurkat Whole Cell Lysate, Lane B: U87MG Whole Cell Lysate. Samples (30 µg per lane) were incubated with polyclonal antibody (Product # PA5-117919) with a dilution of 1:500 , followed by Goat Anti-Rabbit IgG (H+L)/HRP using a dilution of 1:10,000. Assay was performed under reducing conditions. Predicted band size: 62 kDa, Observed band size: 62 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using CRMP2 Polyclonal Antibody (Product # PA5-117919) at 1:500 dilution. Lane A: Jurkat Whole Cell Lysate, Lane B: U87MG Whole Cell Lysate. Lysates/proteins at 30 μg per lane. Secondary antibody: Goat Anti-Rabbit IgG (H+L)/HRP at 1:10,000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size: 62 kDa. Observed band size: 62 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot of CRMP2 in Lane A: Jurkat Whole Cell Lysate, Lane B: U87MG Whole Cell Lysate. Samples (30 µg per lane) were incubated with polyclonal antibody (Product # PA5-117919) with a dilution of 1:500 , followed by Goat Anti-Rabbit IgG (H+L)/HRP using a dilution of 1:10,000. Assay was performed under reducing conditions. Predicted band size: 62 kDa, Observed band size: 62 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using CRMP2 Polyclonal Antibody (Product # PA5-117919) at 1:500 dilution. Lane A: Jurkat Whole Cell Lysate, Lane B: U87MG Whole Cell Lysate. Lysates/proteins at 30 μg per lane. Secondary antibody: Goat Anti-Rabbit IgG (H+L)/HRP at 1:10,000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size: 62 kDa. Observed band size: 62 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CRMP2 Polyclonal Antibody (Product # PA5-117919) and 58, 62 kDa bands corresponding to DPYSL2 was observed across neuronal cell lines and MCF 10A except MCF7, SK-BR-3 which are low expressing models for CRMP2. Relative expression was also observed in brain tissues when compared to Mouse Brown Adipose and Mouse Colon. Whole cell extracts (30 µg lysate) of PC-12 (Lane 1), Neuro-2a (Lane 2), SH-SY5Y (Lane 3), U-87 MG (Lane 4), MCF7 (Lane 5), MCF 10A (Lane 6), SK-BR-3 (Lane 7), Mouse Brain (Lane 8), Rat Brain (Lane 9), Mouse Brown Adipose (Lane 10), Mouse Colon (Lane 11) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of CRMP2 in U2OS cells. Cells were fixed with 4% PFA, permeabilzed with 0.1% Triton X-100 in PBS, blocked with 10% serum, and incubated with CRMP2 Polyclonal Antibody (Product # PA5-117919, 1:200) at 4°C overnight. Then cells were stained with the Alexa Fluor®594-conjugated Goat Anti-rabbit IgG secondary antibody (red) and counterstained with DAPI (blue). Positive staining was localized to cytoplasm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of DPYSL2 was performed using 70% confluent log phase SH-SY5Y and MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CRMP2 Polyclonal Antibody (Product # PA5-117919) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents MCF7 cells with low expression of CRMP2 protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: CRMP2 Immunoprecipitation using: Lane A: 0.5 mg Jurkat Whole Cell Lysate 4 µL with CRMP2 Polyclonal Antibody (Product # PA5-117919) and 60 μg of Immunomagnetic beads Protein A/G. Primary antibody: CRMP2 Polyclonal Antibody, at 1:100 dilution. Secondary antibody: Goat Anti-Rabbit IgG (H+L) /HRP at 1:10,000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size: 62 kDa. Observed band size: 62 kDa.

PA5-117919