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RanGAP1 Recombinant Rabbit Monoclonal Antibody [JG37-41]

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Specification

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Catalog# ET7108-48

RanGAP1 Recombinant Rabbit Monoclonal Antibody [JG37-41]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

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

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

AB_3070846

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:100

  • IF-Tissue

  • 1:100

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:100

  • IP

  • 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.

UNIPROT

SwissProt: P46060 Human

SwissProt: P46061 Mouse

Entrez Gene: 362965 Rat

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

Expand

Images

  • Western blot analysis of RanGAP1 on different lysates with Rabbit anti-RanGAP1 antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: SH-SY5Y cell lysate<br />Lane 5: Jurkat cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 63 kDa<br />Observed band size: 63/80 kDa<br /><br />Exposure time: 3 minutes 10 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) 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 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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) 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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) 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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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 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.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7108-48" style="font-weight: bold;text-decoration: underline;">ET7108-48</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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).

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