HA721177

MCM2 Recombinant Rabbit Monoclonal Antibody [PD00-90]

cat.: HA721177

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PD00-90

Form:	Liquid
Storage condition:	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:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	0.9415ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 102 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within
Positive control:	HL-60 cell lysate, SiHa cell lysate, human tonsil tissue, human cervical carcinoma tissue, human ovarian carcinoma tissue, human small intestine tissue, LoVo, HL-60.
Subcellular location:	Nucleus, Chromosome.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:2,000 1:800-1:2,000 1:100 1:500-1:1,000
Uniprot #:	SwissProt: P49736 Human
Alternative names:	BM28 CCNL 1 CCNL1 CDC like 1 cdc19 CDCL 1 CDCL1 Cell devision cycle like 1 Cyclin like 1 D3S3194 DNA replication licensing factor MCM2 KIAA0030 MCM 2 MCM2 MCM2 minichromosome maintenance deficient 2 mitotin MCM2 minichromosome maintenance deficient 2 mitotin (S. cerevisiae) MCM2 minichromosome maintenance deficient 2, mitotin MCM2_HUMAN MGC10606 Minichromosome maintenance complex component 2 Minichromosome maintenance deficient 2 (mitotin) Minichromosome maintenance deficient 2 mitotin Minichromosome maintenance protein 2 Minichromosome maintenance protein 2 homolog Mitotin Nuclear protein BM28 OTTHUMP00000216047 OTTHUMP00000216050

Images

	Fig1: Western blot analysis of MCM2 on different lysates with Rabbit anti-MCM2 antibody (HA721177) at 1/2,000 dilution. Lane 1: HL-60 cell lysate Lane 2: SiHa cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 102 kDa Observed band size: 130 kDa Exposure time: 1 minute; 8% 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 (HA721177) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
	Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA721177) 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/800 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 (HA721177) at 1/800 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.

	Fig4: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-MCM2 antibody (HA721177) 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 (HA721177) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/2,000 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 (HA721177) at 1/2,000 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: Immunocytochemistry analysis of LoVo cells labeling MCM2 with Rabbit anti-MCM2 antibody (HA721177) 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-MCM2 antibody (HA721177) 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.

Fig7: Flow cytometric analysis of HL-60 cells labeling MCM2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721177, 1ug/ml) (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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