Ras Recombinant Rabbit Monoclonal Antibody [JF10-11]
Catalog# ET1702-94
Ras Recombinant Rabbit Monoclonal Antibody [JF10-11]
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WB
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IF-Cell
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IP
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FC
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Human
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Mouse
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Rat
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Zebrafish
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unconjugated
Overview
Product Name
Ras Recombinant Rabbit Monoclonal Antibody [JF10-11]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Ras aa 21-64 / 189.
Species Reactivity
Human, Mouse, Rat, Zebrafish
Validated Applications
WB, IF-Cell, IP, FC
Target Molecular Weight
Predicted band size: 21 kDa
Positive Control
MCF7 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, Mouse brain tissue lysate, Mouse ovary tissue lysate, Rat brain tissue lysate, 293T cell lysate, Zebrafish tissue lysate, HeLa, PC-12.
Conjugation
unconjugated
Clone Number
JF10-11
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
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100-1:500
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IP
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Use at an assay dependent concentration.
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FC
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1:1,000
Target
Function
The mammalian c-H-, c-K- and N-Ras proto-oncogenes encode guanine nucleotide-binding proteins that are ubiquitously expressed in vertebrate cells. c-H- and c-K-Ras are cellular homologs of the v-H and v-K-Ras sequences originally isolated from the Harvey and Kirsten strains of rat sarcoma virus. Ras-encoded proteins bind GDP and GTP with high affinity and possess a low level intrinsic GTPase activity that can be stimulated over 100-fold by interaction with cytosolic GTPase activating protein (GAP), a potential effector for Ras p21 function. Point mutations at amino acids 12, 13, 59 and 61 within domains responsible for GTP binding and hydrolysis activate Ras proteins to their oncogenic form and block the ability of the GTPase activity to be stimulated by GAP. Several additional proteins with GAP activity have been identified and shown to interact with p21 Ras or other members of the Ras gene family.
Background References
1. Zhou M et al. VPS35 binds farnesylated N-Ras in the cytosol to regulate N-Ras trafficking. J Cell Biol 214:445-58 (2016).
2. Meng X et al. RPL23 Links Oncogenic RAS Signaling to p53-Mediated Tumor Suppression. Cancer Res 76:5030-9 (2016).
Sequence Similarity
Belongs to the small GTPase superfamily. Ras family.
Post-translational Modification
Palmitoylated by the ZDHHC9-GOLGA7 complex. Depalmitoylated by ABHD17A, ABHD17B and ABHD17C. A continuous cycle of de- and re-palmitoylation regulates rapid exchange between plasma membrane and Golgi.; Acetylation at Lys-104 prevents interaction with guanine nucleotide exchange factors (GEFs).; Ubiquitinated by the BCR(LZTR1) E3 ubiquitin ligase complex at Lys-170 in a non-degradative manner, leading to inhibit Ras signaling by decreasing Ras association with membranes.; Phosphorylation at Ser-89 by STK19 enhances NRAS-association with its downstream effectors.
Subcellular Location
Cytoplasm, Cell membrane, Golgi apparatus, Nucleus.
UNIPROT
Synonyms
C-BAS/HAS antibody
c-H-ras antibody
C-HA-RAS1 antibody
CTLO antibody
GTPase HRas antibody
GTPase KRas antibody
GTPase NRas antibody
H-Ras-1 antibody
H-RASIDX antibody
Ha-Ras antibody
ExpandImages
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Western blot analysis of Ras on different lysates with Rabbit anti-Ras antibody (ET1702-94) at 1/1,000 dilution.
Lane 1: MCF7 cell lysate (15 µg/Lane)
Lane 2: HeLa cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (30 µg/Lane)
Lane 5: Mouse ovary tissue lysate (30 µg/Lane)
Lane 6: Rat brain tissue lysate (30 µg/Lane)
Predicted band size: 21 kDa
Observed band size: 21 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 (ET1702-94) 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. -
Immunocytochemistry analysis of HeLa cells labeling Ras with Rabbit anti-Ras antibody (ET1702-94) 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-Ras antibody (ET1702-94) 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 Ras with Rabbit anti-Ras antibody (ET1702-94) 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-Ras antibody (ET1702-94) 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 HeLa cells labeling Ras.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-94, 1/1,000) (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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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