ET1602-5

M6PR Recombinant Rabbit Monoclonal Antibody [SR45-09]

cat.: ET1602-5

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SR45-09

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 274 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human M6PR aa 2,442-2,491 / 2,491.
Positive control:	HeLa cell lysate, SK-OV-3 cell lysate, SiHa cell lysate, SK-MEL-28 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, PC-12 cell lysate, HeLa, NIH/3T3, C6, human liver tissue, human tonsil tissue, mouse colon tissue, rat colon tissue.
Subcellular location:	Golgi apparatus membrane, Endosome membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:2,000-1:10,000 1:100-1:250 1:50-1:200 1:200 1:1,000 Use at an assay dependent concentration.
Uniprot #:	SwissProt: P11717 Human \| Q07113 Mouse Entrez Gene: 25151 Rat
Alternative names:	300 kDa mannose 6 phosphate receptor 300 kDa mannose 6-phosphate receptor Cation independent mannose 6 phosphate receptor Cation-independent mannose-6-phosphate receptor CD222 CD222 antigen CI Man 6 P receptor CI Man-6-P receptor CI MPR CI-M6PR CI-MPR CIMPR IGF 2 receptor IGF 2R IGF II receptor IGF-II receptor IGF2 receptor Igf2r Insulin like growth factor 2 receptor Insulin like growth factor II receptor Insulin-like growth factor 2 receptor Insulin-like growth factor II receptor M6P R M6P/IGF2 receptor M6P/IGF2R M6PR mannose 6 phosphate receptor mannose 6 phosphate receptor, cation independent MPR 300 MPR300 MPRI MPRI_HUMAN

Images

Fig1: Western blot analysis of M6PR on different lysates with Rabbit anti-M6PR antibody (ET1602-5) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: SK-OV-3 cell lysate
Lane 3: SiHa cell lysate
Lane 4: SK-MEL-28 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 274 kDa
Observed band size: 274 kDa

Exposure time: 6 seconds; ECL: K1801;

3-8% 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 (ET1602-5) at 1/5,000 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 M6PR on different lysates with Rabbit anti-M6PR antibody (ET1602-5) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si M6PR cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 274 kDa
Observed band size: 274 kDa

Exposure time: 21 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 (ET1602-5) 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.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling M6PR with Rabbit anti-M6PR antibody (ET1602-5) 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-M6PR antibody (ET1602-5) 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: Immunocytochemistry analysis of NIH/3T3 cells labeling M6PR with Rabbit anti-M6PR antibody (ET1602-5) 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-M6PR antibody (ET1602-5) 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 (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.
	Fig5: Immunocytochemistry analysis of C6 cells labeling M6PR with Rabbit anti-M6PR antibody (ET1602-5) at 1/250 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-M6PR antibody (ET1602-5) at 1/250 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-M6PR antibody (ET1602-5) at 1/200 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 (ET1602-5) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-M6PR antibody (ET1602-5) at 1/200 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 (ET1602-5) 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-M6PR antibody (ET1602-5) at 1/200 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 (ET1602-5) 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.

Fig9: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-M6PR antibody (ET1602-5) at 1/200 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 (ET1602-5) 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.

Fig10: Flow cytometric analysis of HeLa cells labeling M6PR.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1602-5, 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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