EM1901-83

DNMT1 Mouse Monoclonal Antibody [A3A8]

cat.: EM1901-83

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	A3A8

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein G affinity purified.
Molecular weight:	Predicted band size: 183 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within Human Dnmt1 aa 500-600 / 1616.
Positive control:	HeLa cell lysate, 293T cell lysate, HepG2 cell lysate, Jurkat cell lysate, SH-SY5Y cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, mouse ovary tissue lysate, mouse testis tissue lysate, rat ovary tissue lysate, rat testis tissue lysate, HeLa, human testis tissue, human tonsil tissue, mouse ovary tissue, mouse testis tissue, rat ovary tissue, rat testis tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IHC-P IF-Cell	1:1,000-1:4,000 1:200-1:1,000 1:400
Uniprot #:	SwissProt: P26358 Human \| P13864 Mouse \| Q9Z330 Rat
Alternative names:	ADCADN AIM CXXC finger protein 9 CXXC-type zinc finger protein 9 CXXC9 DNA (cytosine 5 ) methyltransferase 1 DNA (cytosine-5)-methyltransferase 1 DNA methyltransferase 1 DNA methyltransferase HsaI DNA methyltransferase M.HsaI. DNA MTase DNA MTase HsaI DNMT 1 DNMT Dnmt1 DNMT1_HUMAN Dnmt1o FLJ16293 HSN1E M.HsaI MCMT Met1 MGC104992 mMmul MommeD2

Images

Fig1: Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (EM1901-83) at 1/4,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: SH-SY5Y cell lysate
Lane 6: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 183 kDa
Observed band size: 183 kDa

Exposure time: 3 minutes; 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 (EM1901-83) at 1/4,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of DNMT1 on different lysates with Mouse anti-DNMT1 antibody (EM1901-83) at 1/1,000 dilution.

Lane 1: 293T cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: Mouse ovary tissue lysate (40 µg/Lane)
Lane 4: Mouse testis tissue lysate (40 µg/Lane)
Lane 5: Rat ovary tissue lysate (40 µg/Lane)
Lane 6: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 183 kDa
Observed band size: 183 kDa

Exposure time: 3 minutes; 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 (EM1901-83) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling DNMT1 with Mouse anti-DNMT1 antibody (EM1901-83) at 1/400 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 Mouse anti-DNMT1 antibody (EM1901-83) at 1/400 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) 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 (EM1901-83) 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 tonsil tissue with Mouse anti-DNMT1 antibody (EM1901-83) at 1/200 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 (EM1901-83) at 1/200 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 mouse ovary tissue with Mouse anti-DNMT1 antibody (EM1901-83) 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 (EM1901-83) 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 testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) 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 (EM1901-83) 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 rat ovary tissue with Mouse anti-DNMT1 antibody (EM1901-83) 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 (EM1901-83) 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 testis tissue with Mouse anti-DNMT1 antibody (EM1901-83) 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 (EM1901-83) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.