PHGDH Recombinant Mouse Monoclonal Antibody [A10H9-R]
cat.: HA601349
Product Type: Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: A10H9-R
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 57 kDa
Isotype: IgG1
Immunogen: Recombinant protein within human PHGDH aa 1-533.
Positive control: HeLa cell lysate, HEK-293 cell lysate, Jurkat cell lysate, MCF7 cell lysate, A549 cell lysate, HepG2 cell lysate, MDA-MB-468 cell lysate, SK-MEL-28 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, RAW264.7 cell lysate, C6 cell lysate, mouse pancreas tissue lysate, rat pancreas tissue lysate, HeLa, NIH/3T3, human breast cancer tissue, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location: Cytosol, extracellular exosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:2,000
1:100
1:1,000
Uniprot #: SwissProt: O43175 Human | Q61753 Mouse | O08651 Rat
Alternative names: 3 PGDH 3-PGDH 3-phosphoglycerate dehydrogenase 3PGDH D-3-phosphoglycerate dehydrogenase EC 1.1.1.95 Epididymis secretory protein Li 113 HEL S 113 NLS NLS1 PDG PGAD PGD PGDH PGDH3 Phgdh PHGDHD Phosphoglycerate dehydrogenase SERA SERA_HUMAN
Images
HA601349_1.jpg Fig1: Western blot analysis of PHGDH on different lysates with Mouse anti-PHGDH antibody (HA601349) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HEK-293 cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: A549 cell lysate (20 µg/Lane)
Lane 6: HepG2 cell lysate (20 µg/Lane)
Lane 7: MDA-MB-468 cell lysate (20 µg/Lane)
Lane 8: SK-MEL-28 cell lysate (20 µg/Lane)
Lane 9: NIH/3T3 cell lysate (20 µg/Lane)

Predicted band size: 57 kDa
Observed band size: 55 kDa

Exposure time: 2 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 (HA601349) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.
HA601349_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PHGDH with Mouse anti-PHGDH antibody (HA601349) 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 Mouse anti-PHGDH antibody (HA601349) at 1/100 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) was used as the secondary antibody at 1/1,000 dilution.
HA601349_3.jpg Fig3: Western blot analysis of PHGDH on different lysates with Mouse anti-PHGDH antibody (HA601349) at 1/2,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: Neuro-2a cell lysate (20 µg/Lane)
Lane 3: RAW264.7 cell lysate (20 µg/Lane)
Lane 4: C6 cell lysate (20 µg/Lane)
Lane 5: Mouse pancreas tissue lysate (40 µg/Lane)
Lane 6: Rat pancreas tissue lysate (40 µg/Lane)

Predicted band size: 57 kDa
Observed band size: 55 kDa

Exposure time: 10 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 (HA601349) at 1/2,000 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.
HA601349_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling PHGDH with Mouse anti-PHGDH antibody (HA601349) 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 Mouse anti-PHGDH antibody (HA601349) at 1/100 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) was used as the secondary antibody at 1/1,000 dilution.
HA601349_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-PHGDH antibody (HA601349) at 1/1,000 dilution and competitor's antibody at 1/100 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 (HA601349) at 1/100 dilution and competitor's antibody 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.
HA601349_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-PHGDH antibody (HA601349) at 1/1,000 dilution and competitor's antibody at 1/100 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 (HA601349) at 1/100 dilution and competitor's antibody 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.
HA601349_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Mouse anti-PHGDH antibody (HA601349) at 1/1,000 dilution and competitor's antibody at 1/100 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 (HA601349) at 1/100 dilution and competitor's antibody 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.
HA601349_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-PHGDH antibody (HA601349) at 1/1,000 dilution and competitor's antibody at 1/100 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 (HA601349) at 1/100 dilution and competitor's antibody 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.
HA601349_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue (negative) with Mouse anti-PHGDH antibody (HA601349) at 1/1,000 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 (HA601349) 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.
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