PKM2 Recombinant Rabbit Monoclonal Antibody [PSH11-74]
cat.: HA751429
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH11-74
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 58 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human PKM2 aa 381-430.
Positive control: HeLa cell lysate, MCF7 cell lysate, A549 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, COS-1 cell lysate, HeLa, NIH/3T3, C6, human breast cancer tissue, human colon cancer tissue, human liver tissue, human testis tissue, mouse liver tissue, mouse testis tissue, rat liver tissue, rat testis tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:10,000
1:100
1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P14618-1 Human | P52480-1 Mouse | P11980-2 Rat
Alternative names: CTHBP Cytosolic thyroid hormone-binding protein KPYM_HUMAN OIP-3 Opa-interacting protein 3 p58 pkm PKM1 PKM2 Pyruvate kinase 2/3 Pyruvate kinase muscle isozyme Pyruvate kinase PKM THBP1 Thyroid hormone-binding protein 1 Tumor M2-PK
Images
HA751429_1.jpg Fig1: Western blot analysis of PKM2 on different lysates with Rabbit anti-PKM2 antibody (HA751429) at 1/10,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
Lane 5: C6 cell lysate (15 µg/Lane)
Lane 6: Mouse skeletal muscle tissue lysate (negative) (20 µg/Lane)
Lane 7: COS-1 cell lysate (15 µg/Lane)

Predicted band size: 58 kDa
Observed band size: 58 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 (HA751429) at 1/10,000 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.
HA751429_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PKM2 with Rabbit anti-PKM2 antibody (HA751429) 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-PKM2 antibody (HA751429) 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 (HA601187, 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.
HA751429_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling PKM2 with Rabbit anti-PKM2 antibody (HA751429) 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-PKM2 antibody (HA751429) 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 (HA601187, 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.
HA751429_4.jpg Fig4: Immunocytochemistry analysis of C6 cells labeling PKM2 with Rabbit anti-PKM2 antibody (HA751429) 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-PKM2 antibody (HA751429) 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 (HA601187, 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.
HA751429_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue (negative) with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue (negative) with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_13.jpg Fig13: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_14.jpg Fig14: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_15.jpg Fig15: Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue (negative) with Rabbit anti-PKM2 antibody (HA751429) 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 (HA751429) 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.
HA751429_16.jpg Fig16: Flow cytometric analysis of HeLa cells labeling PKM2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751429, 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).
HA751429_17.jpg Fig17: Flow cytometric analysis of NIH/3T3 cells labeling PKM2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751429, 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).
HA751429_18.jpg Fig18: Flow cytometric analysis of C6 cells labeling PKM2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751429, 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).
HA751429_19.jpg Fig19: Western blot analysis of PKM2 on different lysates with Rabbit anti-PKM2 antibody (HA751429) at 1/2,000 dilution.

Lane 1: Human PKM1 recombinant protein (50 ng/Lane)
Lane 2: Human PKM2 recombinant protein (50 ng/Lane)

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 (HA751429) 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.
HA751429_20.jpg Fig20: PKM2 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751429 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751429 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751429 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751429 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 12 seconds; ECL: K1801
HA751429_21.jpg Fig21: PKM2 was immunoprecipitated from 0.2 mg RAW264.7 cell lysate with HA751429 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751429 at 1/10,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: RAW264.7 cell lysate (input)
Lane 2: HA751429 IP in RAW264.7 cell lysate
Lane 3: Rabbit IgG instead of HA751429 in RAW264.7 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 25 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.