Nogo Recombinant Rabbit Monoclonal Antibody [JM02-34]
cat.: ET1705-63
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JM02-34
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 130 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Nogo aa 111-160 / 1192.
Positive control: Mouse neuron, HUVEC cell lysate, HepG2 cell lysate, A549 cell lysate, mouse skeletal muscle tissue lysates, human breast tissue, human fetal skeletal muscle tissue, mouse testis tissue, mouse brain tissue, A549.
Subcellular location: Endoplasmic reticulum membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:2,000
1:50-1:200
1:50-1:200
1:200-1:1,000
1:1,000
1:10-1:50
Uniprot #: SwissProt: Q9NQC3 Human | Q99P72 Mouse
Alternative names: 1110020G17Rik AA407876 AA409940 AA960376 ASY C130026I10Rik Foocen Glut4 vesicle 20 kDa protein Human NogoA Kiaa0886 KIAA4153 MGC116054 MGC139261 mKIAA0886 mKIAA4153 My043 protein Nbla00271 Nbla10545 Neurite growth inhibitor 220 Neurite Growth Inhibitor 220, included Neurite outgrowth inhibitor Neuroendocrine-specific protein Neuroendocrine-specific protein C homolog NI-250 NI220/250 Nogo A NOGO Nogo B Nogo C Nogo protein NOGOC NSP NSP-CL rat N Reticulon 4 Reticulon 5 Reticulon-4 Reticulon-5 RTN X RTN-x Rtn4 Rtn4 reticulon 4 RTN4-A RTN4-B1 RTN4-B2 RTN4-C RTN4_HUMAN Vp20
Images
ET1705-63_1.jpg Fig1: Immunocytochemistry analysis of mouse neuron cells labeling Nogo with Rabbit anti-Nogo antibody (ET1705-63) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nogo antibody (ET1705-63) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1705-63_2.jpg Fig2: Western blot analysis of Nogo on different lysates with Rabbit anti-Nogo antibody (ET1705-63) at 1/1,000 dilution.

Lane 1: HUVEC cell lysate
Lane 2: HepG2 cell lysate
Lane 3: A549 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 40-50 kDa

Exposure time: 6 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-63) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1705-63_3.jpg Fig3: Western blot analysis of Nogo on mouse skeletal muscle tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-63, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1705-63_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Nogo antibody (ET1705-63) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-63) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-63_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human fetal skeletal muscle tissue using anti-Nogo antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-63, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-63_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Nogo antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-63, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-63_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Nogo antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-63, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-63_8.jpg Fig8: Immunocytochemistry analysis of A549 cells labeling Nogo with Rabbit anti-Nogo antibody (ET1705-63) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nogo antibody (ET1705-63) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1705-63_9.jpg Fig9: Flow cytometric analysis of A549 cells labeling Nogo.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-63, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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