GRASP65 Recombinant Rabbit Monoclonal Antibody [JB36-34]
Catalog# ET7107-13
GRASP65 Recombinant Rabbit Monoclonal Antibody [JB36-34]
-
WB
-
IF-Cell
-
IF-Tissue
-
IHC-P
-
FC
-
IP
-
Human
-
unconjugated
Overview
Product Name
GRASP65 Recombinant Rabbit Monoclonal Antibody [JB36-34]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human GRASP65 aa 396-440 / 440.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 46 kDa
Positive Control
SK-Br-3 cell lysate, HepG2 cell lysate, 293T cell lysate, HeLa cell lysate, HepG2, human uterus tissue, human colon carcinoma tissue, MCF-7.
Conjugation
unconjugated
Clone Number
JB36-34
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000
-
IF-Cell
-
1:50-1:200
-
IHC-P
-
1:50-1:200
-
FC
-
1:50-1:100
-
IP
-
Use at an assay dependent concentration.
Target
Function
Golgi reassembly-stacking protein of 65 kDa (GRASP65) also known as Golgi reassembly-stacking protein 1 (GORASP1) is a protein that in humans is encoded by the GORASP1 gene. The Golgi complex plays a key role in the sorting and modification of proteins exported from the endoplasmic reticulum. The GRASP65 protein is a peripheral membrane protein anchored to the lipid bilayer through myristoylation of a glycine residue near the protein's amino terminus. It is involved in establishing the stacked structure of the Golgi apparatus and linking the stacks into larger ribbons in vertebrate cells. It is a caspase-3 substrate, and cleavage of this encoded protein contributes to Golgi fragmentation in apoptosis. GRASP65 can form a complex with the Golgi matrix protein GM130, and this complex binds to the vesicle docking protein p115. Several alternatively spliced transcript variants of this gene have been identified, but their full-length natures have not been determined.
Background References
1. Puthenveedu M.A. et al. GM130 and GRASP65-dependent lateral cisternal fusion allows uniform Golgi-enzyme distribution. Nat. Cell Biol. 8:238-248(2006).
2. Marra P. et al. The GM130 and GRASP65 Golgi proteins cycle through and define a subdomain of the intermediate compartment. Nat. Cell Biol. 3:1101-1113(2001).
Sequence Similarity
Belongs to the GORASP family.
Post-translational Modification
Phosphorylated by CDC2/B1 and PLK kinases during mitosis. Phosphorylation cycle correlates with the cisternal stacking cycle. Phosphorylation of the homodimer prevents the association of dimers into higher-order oligomers, leading to cisternal unstacking.; Target for caspase-3 cleavage during apoptosis. The cleavage contributes to Golgi fragmentation and occurs very early in the execution phase of apoptosis.; Myristoylated.
Subcellular Location
Golgi apparatus.
UNIPROT
Synonyms
FLJ23443 antibody
Golgi peripheral membrane protein p65 antibody
Golgi phosphoprotein 5 antibody
Golgi reassembly and stacking protein 1 antibody
Golgi reassembly and stacking protein 65 kDa antibody
Golgi reassembly and stacking protein, 65-kD antibody
Golgi reassembly stacking protein 1 65kDa antibody
Golgi reassembly stacking protein 1 antibody
Golgi reassembly stacking protein of 65 kDa antibody
Golgi reassembly-stacking protein 1 antibody
ExpandImages
-
Western blot analysis of GRASP65 on different lysates with Rabbit anti-GRASP65 antibody (ET7107-13) at 1/1,000 dilution.
Lane 1: SK-Br-3 cell lysate
Lane 2: HepG2 cell lysate
Lane 3: 293T cell lysate
Lane 4: HeLa cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 46 kDa
Observed band size: 70 kDa
Exposure time: 25 seconds; ECL: K1801;
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 (ET7107-13) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
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. -
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. -
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.
-
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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"