ET7106-97

PFKM Recombinant Rabbit Monoclonal Antibody [JU53-31]

cat.: ET7106-97

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JU53-31

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: 85 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human PFKM aa 107-156 / 780.
Positive control:	HeLa cell lysate, Ramos cell lysate, mouse heart tissue lysate, mouse skeletal muscle tissue lysate, rat heart tissue lysate, rat skeletal muscle tissue lysate, A549 cell lysate, LOVO, PC-3M, SH-SY5Y, rat brain tissue, human liver carcinoma tissue, mouse skeletal muscle tissue, human thyroid tissue, human kidney tissue.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P08237 Human \| P47857 Mouse \| P47858 Rat
Alternative names:	6 Phosphofructokinase Muscle Type 6-phosphofructokinase 6-phosphofructokinase muscle type EC 2.7.1.1 EC 2.7.1.11 GSD7 K6PF_HUMAN MGC8699 muscle type PFK, muscle type PFK-A PFKA PFKM PFKX Phosphofructo 1 Kinase Isozyme A Phosphofructo-1-kinase isozyme A Phosphofructokinase 1 Phosphofructokinase M Phosphofructokinase, muscle Phosphofructokinase, muscle type Phosphofructokinase, polypeptide X Phosphofructokinase-M Phosphohexokinase

Images

Fig1: Western blot analysis of PFKM on different lysates with Rabbit anti-PFKM antibody (ET7106-97) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Ramos cell lysate
Lane 3: Mouse heart tissue lysate
Lane 4: Mouse skeletal muscle tissue lysate
Lane 5: Rat heart tissue lysate
Lane 6: Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 75 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 (ET7106-97) 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.

Fig2: Western blot analysis of PFKM on different lysates with Rabbit anti-PFKM antibody (ET7106-97) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD PFKM cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 75 kDa

Exposure time: 3 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 (ET7106-97) 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: ICC staining of PFKM in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: ICC staining of PFKM in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: ICC staining of PFKM in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-97, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-PFKM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7106-97, 1/50) for 30 minutes 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 liver carcinoma tissue using anti-PFKM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7106-97, 1/50) for 30 minutes 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 skeletal muscle tissue using anti-PFKM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7106-97, 1/50) for 30 minutes 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 human thyroid tissue using anti-PFKM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7106-97, 1/50) for 30 minutes 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: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PFKM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7106-97, 1/50) for 30 minutes 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.

Fig11: Flow cytometric analysis of PFKM was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7106-97, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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