Phospho-MCM2 (S41) Recombinant Rabbit Monoclonal Antibody [JE50-04]
cat.: HA722279
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: JE50-04
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: 102 kDa
Isotype: IgG
Immunogen: Synthetic peptide corresponding to residues surrounding Ser41 of human MCM2 protein.
Positive control: A431 cell lysate, HeLa cell lysate, HEK-293 cell lysate, THP-1 cell lysate, mouse spleen tissue lysate, mouse thymus tissue lysate, rat spleen tissue lysate, rat thymus tissue lysate, THP-1, human tonsil tissue, human spleen tissue, mouse spleen tissue, rat spleen tissue.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:1,000
1:500
1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P49736 Human | P97310 Mouse
Entrez Gene: 312538 Rat
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
HA722279_1.jpg Fig1: Western blot analysis of Phospho-MCM2 (S41) on different lysates with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) at 1/1,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: HEK-293 cell lysate (20 µg/Lane)
Lane 4: THP-1 cell lysate (20 µg/Lane)
Lane 5: Mouse spleen tissue lysate (30 µg/Lane)
Lane 6: Mouse thymus tissue lysate (30 µg/Lane)
Lane 7: Rat spleen tissue lysate (30 µg/Lane)
Lane 8: Rat thymus tissue lysate (30 µg/Lane)

Predicted band size: 102 kDa
Observed band size: 130 kDa

Exposure time: Lane 1-4: 8 seconds; Lane 5-8: 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 (HA722279) 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.
HA722279_2.jpg Fig2: Immunocytochemistry analysis of THP-1 cells labeling Phospho-MCM2 (S41) with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Phospho-MCM2 (S41) antibody (HA722279) at 1/500 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 (M1305-2, 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.
HA722279_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human tonsil tissue untreated / treated with λpp with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) 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 (HA722279) 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.
HA722279_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) 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 (HA722279) 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.
HA722279_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) 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 (HA722279) 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.
HA722279_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-MCM2 (S41) antibody (HA722279) 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 (HA722279) 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.
HA722279_7.jpg Fig7: Flow cytometric analysis of THP-1 cells labeling Phospho-MCM2 (S41).

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722279, 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).
HA722279_8.jpg Fig8: Phospho-MCM2 (S41) was immunoprecipitated from 0.2 mg THP-1 cell lysate with HA722279 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722279 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 cell lysate (input)
Lane 2: HA722279 IP in THP-1 cell lysate
Lane 3: Rabbit IgG instead of HA722279 in THP-1 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 2 minutes 5 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.