RanGAP1 Recombinant Rabbit Monoclonal Antibody [JG37-41]
Catalog# ET7108-48
RanGAP1 Recombinant Rabbit Monoclonal Antibody [JG37-41]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
RanGAP1 Recombinant Rabbit Monoclonal Antibody [JG37-41]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human RanGAP1 aa 1-50 / 587.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 63 kDa
Positive Control
HeLa cell lysate, A431 cell lysate, MCF7 cell lysate, SH-SY5Y cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human testis tissue, mouse testis tissue, rat testis tissue, MCF7, NIH/3T3, PC-12.
Conjugation
unconjugated
Clone Number
JG37-41
RRID
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC | IF-Cell | |
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| human |
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| mouse |
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| rat |
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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
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WB
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1:1,000
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IF-Cell
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1:100
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IF-Tissue
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1:100
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IHC-P
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1:50-1:200
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FC
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1:100
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IP
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1:10-1:50
Target
Function
The small Ras related protein Ran, also called TC4, is a nuclear localized GTPase implicated in a diverse array of cellular processes including DNA replication, entry into and exit from mitosis and the transport of RNA and proteins through the nuclear pore complex. Like Ras, active Ran GTP and inactive Ran GDP levels are tightly regulated by guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). The abundant GEF RCC1 (regulator of chromosome condensation 1) increases the rate at which Ran exchanges GDP for GTP. Ran GAP1 opposes the effects of RCC1 by increasing the rate at which Ran hydrolyzes GTP to GDP. A protein designated Ran BP1 has no intrinsic GAP activity and functions as a GEF inhibitor deactivating RCC1 and thereby indirectly increasing the ratio of Ran GDP to Ran GTP. Ran BP2 has been proposed as the Ran GTP docking site at the periphery of the nuclear pore complex.
Background References
1. Bischoff F R et al. RanGAP1 induces GTPase activity of nuclear Ras-related Ran. Proc Natl Acad Sci USA 91:2587-2591 (1994) .
2. Joseph J et al. SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles. J Cell Biol 156:595-602 (2002) .
Sequence Similarity
Belongs to the RNA1 family.
Tissue Specificity
Highly expressed in brain, thymus and testis.
Post-translational Modification
Phosphorylation occurs before nuclear envelope breakdown and continues throughout mitosis. Phosphorylated by the M-phase kinase cyclin B/Cdk1, in vitro. Differential timimg of dephosphorylation occurs during phases of mitosis. The phosphorylated form remains associated with RANBP2/NUP358 and the SUMO E2-conjugating enzyme, UBE2I, on nuclear pore complex (NPC) diassembly and during mitosis.; Sumoylated. Sumoylation is necessary for targeting to the nuclear envelope (NE), and for association with mitotic spindles and kinetochores during mitosis. Also required for interaction with RANBP2 and is mediated by UBE2I.
Subcellular Location
Cytoplasm. Cytoskeleton. Nucleus.
Synonyms
Fug 1 antibody
Fug1 antibody
GTPase-activating protein, RAN, 1 antibody
KIAA1835 antibody
MGC20266 antibody
OTTHUMP00000028918 antibody
OTTHUMP00000198755 antibody
OTTHUMP00000198756 antibody
OTTHUMP00000198757 antibody
OTTHUMP00000198758 antibody
ExpandImages
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Western blot analysis of RanGAP1 on different lysates with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: MCF7 cell lysate
Lane 4: SH-SY5Y cell lysate
Lane 5: Jurkat cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 63 kDa
Observed band size: 63/80 kDa
Exposure time: 3 minutes 10 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 (ET7108-48) 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. -
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/200 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 (ET7108-48) at 1/200 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 mouse testis tissue with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/200 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 (ET7108-48) at 1/200 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 rat testis tissue with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/200 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 (ET7108-48) at 1/200 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. -
Immunocytochemistry analysis of MCF7 cells labeling RanGAP1 with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RanGAP1 antibody (ET7108-48) at 1/100 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 (HA601187, 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. -
Immunocytochemistry analysis of NIH/3T3 cells labeling RanGAP1 with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RanGAP1 antibody (ET7108-48) at 1/100 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 (HA601187, 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. -
Immunocytochemistry analysis of PC-12 cells labeling RanGAP1 with Rabbit anti-RanGAP1 antibody (ET7108-48) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RanGAP1 antibody (ET7108-48) at 1/100 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 (HA601187, 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. -
Flow cytometric analysis of MCF7 cells labeling RanGAP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-48, 1/100) (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). -
Flow cytometric analysis of NIH/3T3 cells labeling RanGAP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-48, 1/100) (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). -
Flow cytometric analysis of PC-12 cells labeling RanGAP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-48, 1/100) (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). -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Bone marrow
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET7108-48, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Testis
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET7108-48, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
Citation
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Hepatitis B virus core protein stabilizes RANGAP1 to upregulate KDM2A and facilitate hepatocarcinogenesis
Journal: Cellular Oncology
DOI:
IF: 6.6
Application: WB
Reactivity: Human
Publish date: 2023 Oct
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"