HA721955

MTAP Recombinant Rabbit Monoclonal Antibody [JE63-74]

cat.: HA721955

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IF-Tissue, FC
Clonality:	Monoclonal
Clone number:	JE63-74

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: 31 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human MTAP aa 184-283 / 283.
Positive control:	HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, NIH/3T3, mouse liver tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue FC	1:1,000 1:100 1:500 1:200 1:1,000
Uniprot #:	SwissProt: Q13126 Human \| Q9CQ65 Mouse Entrez Gene: 298227 Rat
Alternative names:	5' methylthioadenosine phosphorylase 5''-methylthioadenosine phosphorylase BDMF c86fus DMSFH DMSMFH Epididymis luminal protein 249 HEL 249 LGMBF MeSAdo phosphorylase Methylthioadenosine phosphorylase MSAP MTA phosphorylase MTAP MTAP_HUMAN MTAPase S methyl 5 thioadenosine phosphorylase S methyl 5' thioadenosine phosphorylase S-methyl-5''-thioadenosine phosphorylase

Images

Fig1: Western blot analysis of MTAP on different lysates with Rabbit anti-MTAP antibody (HA721955) at 1/2,000 dilution.

Lane 1: HCT 116-si NT cell lysate
Lane 2: HCT 116-si MTAP cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 40 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 (HA721955) at 1/2,000 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 MTAP on different lysates with Rabbit anti-MTAP antibody (HA721955) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 24 seconds;

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 (HA721955) 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.

	Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling MTAP with Rabbit anti-MTAP antibody (HA721955) at 1/100 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-MTAP antibody (HA721955) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-MTAP antibody (HA721955) at 1/500 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 (HA721955) at 1/500 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: Immunofluorescence analysis of paraffin-embedded mouse liver tissue labeling MTAP with Rabbit anti-MTAP antibody (HA721955) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721955, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Fig6: Flow cytometric analysis of NIH/3T3 cells labeling MTAP.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721955, 1μg/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).

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