ET7108-97

Methionine Aminopeptidase 2 / p67 Recombinant Rabbit Monoclonal Antibody [JG39-79]

cat.: ET7108-97

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JG39-79

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 53 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human METAP2 aa 340-478 / 478.
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.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:2,000 1:100 1:200 1:1,000 1:1,000
Uniprot #:	SwissProt: P50579 Human \| O08663 Mouse \| P38062 Rat
Alternative names:	A930035J23Rik AI047573 AL024412 Amp2 AU014659, EIF 2 associated p67 homolog EIF-2-associated p67 Initiation factor 2 associated 67 kDa glycoprotein Initiation factor 2-associated 67 kDa glycoprotein MAP 2 MAP2 MAP2_HUMAN MetAP 2 Metap2 Methionine aminopeptidase 2 Methionyl aminopeptidase 2 MGC102452 MGC127390 MGC53792 MNPEP p67 p67eIF2 Peptidase M Peptidase M2

Images

Fig1: 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.

Fig2: 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.

	Fig3: 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.
	Fig4: 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.
	Fig5: 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.

	Fig6: 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.
	Fig7: 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.
	Fig8: 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.

	Fig9: 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.
	Fig10: 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.
	Fig11: 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).

Fig12: 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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