GRASP65 Recombinant Rabbit Monoclonal Antibody [JB36-34]
cat.: ET7107-13
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: JB36-34
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: 46 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human GRASP65 aa 396-440 / 440.
Positive control: SK-Br-3 cell lysates, HepG2, HUVEC, MCF-7, human colon carcinoma tissue, human uterus tissue.
Subcellular location: Golgi apparatus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:500-1:1,000
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q9BQQ3 Human
Alternative names: FLJ23443 Golgi peripheral membrane protein p65 Golgi phosphoprotein 5 Golgi reassembly and stacking protein 1 Golgi reassembly and stacking protein 65 kDa Golgi reassembly and stacking protein, 65-kD Golgi reassembly stacking protein 1 65kDa Golgi reassembly stacking protein 1 Golgi reassembly stacking protein of 65 kDa Golgi reassembly-stacking protein 1 Golgi reassembly-stacking protein of 65 kDa GOLPH 5 GOLPH5 Gorasp 1 GORASP1 GORS1_HUMAN GRASP 65 GRASP65 MGC118894 MGC118897 P65
Images
ET7107-13_1.jpg Fig1: Western blot analysis of GRASP65 on SK-Br-3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-13, 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.
ET7107-13_2.png Fig2: Immunocytochemistry analysis of HepG2 cells labeling GRASP65 with Rabbit anti-GRASP65 antibody (ET7107-13) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-GRASP65 antibody (ET7107-13) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.
ET7107-13_3.jpg Fig3: ICC staining of GRASP65 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-13, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET7107-13_4.jpg Fig4: ICC staining of GRASP65 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-13, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET7107-13_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-GRASP65 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 (ET7107-13, 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.
ET7107-13_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human uterus tissue with Rabbit anti-GRASP65 antibody (ET7107-13) at 1/100 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 (ET7107-13) at 1/100 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.
ET7107-13_7.jpg Fig7: Flow cytometric analysis of GRASP65 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-13, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.