Hamartin Mouse Monoclonal Antibody [A6F1]
cat.: HA600004
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: A6F1
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 130 kDa
Isotype: IgG1
Immunogen: Recombinant protein within human Hamartin aa 401-600.
Positive control: Hela cell lysate, 293 cell lysate, NIH/3T3 cell lysate, Hela, NIH/3T3, human kidney tissue, human fetal skeletal muscle tissue, PC-3.
Subcellular location: Cytoplasm, Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC

1:500
1:100
1:200-1:600
1:500-1:1,000
Uniprot #: SwissProt: Q92574 Human | Q9EP53 Mouse
Alternative names: Hamartin kiaa0243 LAM TSC Tsc1 Tsc1 gene TSC1_HUMAN Tuberous sclerosis 1 Tuberous sclerosis 1 protein tumor suppressor
Images
HA600004_1.jpg Fig1: Western blot analysis of Hamartin on different lysates with Mouse anti-Hamartin antibody (HA600004) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: 293 cell lysate
Lane 3: NIH/3T3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 145 kDa

Exposure time: 2 minutes;

6% 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 (HA600004) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
HA600004_2.jpg Fig2: Immunocytochemistry analysis of Hela cells labeling Hamartin with Mouse anti-Hamartin antibody (HA600004) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Hamartin antibody (HA600004) at 1/100 dilution in 1% BSA 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.
HA600004_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling Hamartin with Mouse anti-Hamartin antibody (HA600004) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Hamartin antibody (HA600004) at 1/100 dilution in 1% BSA 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.
HA600004_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Hamartin antibody (HA600004) at 1/600 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 (HA600004) at 1/600 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.
HA600004_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human fetal skeletal muscle tissue with Mouse anti-Hamartin antibody (HA600004) 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 (HA600004) 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.
HA600004_6.jpg Fig6: Flow cytometric analysis of PC-3 cells labeling Hamartin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA600004, 1ug/ml) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA600004_7.jpg Fig7: Flow cytometric analysis of NIH/3T3 cells labeling Hamartin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA600004, 1ug/ml) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA600004_8.jpg Fig8: Flow cytometric analysis of Hela cells labeling Hamartin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA600004, 1ug/ml) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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