Methionine Aminopeptidase 2 / p67 Recombinant Rabbit Monoclonal Antibody [JG39-79]
Catalog# ET7108-97
Methionine Aminopeptidase 2 / p67 Recombinant Rabbit Monoclonal Antibody [JG39-79]
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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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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Methionine Aminopeptidase 2 / p67 Recombinant Rabbit Monoclonal Antibody [JG39-79]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human METAP2 aa 340-478 / 478.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Molecular Weight
Predicted band size: 53 kDa
Positive Control
Daudi cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, Rat thymus tissue lysate, K-562, NIH/3T3, human kidney tissue, human testis tissue, mouse kidney tissue, mouse testis tissue, rat kidney tissue, rat testis tissue, NIH3T3.
Conjugation
unconjugated
Clone Number
JG39-79
RRID
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
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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
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IF-Tissue
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1:200
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IHC-P
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1:1,000
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FC
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1:1,000
Target
Function
Methionine aminopeptidases (MetAP), also designated peptidase M proteins, are members of the M24 family of proteins. Both MetAP-1 and MetAP-2 release N-terminal amino acids, usually methionine, from nascent peptides and arylamines. Eukaryotes contain both MetAP-1 and MetAP-2, whereas prokaryotes possess only the MetAP-1 enzyme. MetAP-1 and MetAP-2 control cell proliferation in mammalian cells. MetAP-2 is highly conserved between human and Saccharomyces cerevisiae. Neurofibromin (NF1) regulates MetAp-2 and increased expression of MetAP-2 correlates with several forms of cancer. Inhibitors of MetAP-2 are potential targets in cancer therapeutics, particularly in NF1-associated tumor proliferation. Chemotherapeutic drugs such as ovalicin and fumagillin bind to the active site of and inhibit MetAp-2.
Background References
1. Xiao Q et al. Protein N-terminal processing: substrate specificity of Escherichia coli and human methionine aminopeptidases. Biochemistry 49:5588-5599 (2010).
2. Wang J et al. Physiologically relevant metal cofactor for methionine aminopeptidase-2 is manganese. Biochemistry 42:5035-5042 (2003).
Sequence Similarity
Belongs to the peptidase M24A family. Methionine aminopeptidase eukaryotic type 2 subfamily.
Post-translational Modification
Contains approximately 12 O-linked N-acetylglucosamine (GlcNAc) residues. O-glycosylation is required for EIF2S1 binding.
Subcellular Location
Cytoplasm.
Synonyms
A930035J23Rik antibody
AI047573 antibody
AL024412 antibody
Amp2 antibody
AU014659, antibody
EIF 2 associated p67 homolog antibody
EIF-2-associated p67 antibody
Initiation factor 2 associated 67 kDa glycoprotein antibody
Initiation factor 2-associated 67 kDa glycoprotein antibody
MAP 2 antibody
ExpandImages
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Western blot analysis of Methionine Aminopeptidase 2 / p67 on different lysates with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 dilution.
Lane 1: Daudi cell lysate
Lane 2: K-562 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C2C12 cell lysate
Lane 5: Rat thymus tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 53 kDa
Observed band size: 67 kDa
Exposure time: 30 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 (ET7108-97) 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. -
☑ Knockdown (KD)
Western blot analysis of Methionine Aminopeptidase 2 / p67 on different lysates with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Methionine Aminopeptidase 2 / p67 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 53 kDa
Observed band size: 67 kDa
Exposure time: 6 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 (ET7108-97) at 1/2,000 dilution was used in 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 K-562 cells labeling Methionine Aminopeptidase 2 / p67 with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) 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-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) 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 Methionine Aminopeptidase 2 / p67 with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) 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-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) 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. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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 kidney tissue with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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 kidney tissue with Rabbit anti-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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-Methionine Aminopeptidase 2 / p67 antibody (ET7108-97) at 1/1,000 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 (ET7108-97) at 1/1,000 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. -
Flow cytometric analysis of K-562 cells labeling Methionine Aminopeptidase 2 / p67.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-97, 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). -
Flow cytometric analysis of NIH3T3 cells labeling Methionine Aminopeptidase 2 / p67.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-97, 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).
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