PFKL Monoclonal antibody

PFKL Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human, Mouse, Rat, Rabbit, Pig and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

1H2G2

Cat no : 68385-1-Ig

Synonyms

PFK B, EC:2.7.1.11, ATP-PFK, ATP-dependent 6-phosphofructokinase, liver type, 6-phosphofructokinase type B

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Validation Data Gallery

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  WB analysis using 68385-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68385-1-Ig (PFKL antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

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  IHC staining of human lung cancer using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse liver using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse liver using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovary tumor using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovary tumor using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver using 68385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 68385-1-Ig (PFKL antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HuH-7 using 68385-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed HuH-7 cells using PFKL antibody (68385-1-Ig, Clone: 1H2G2 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of HuH-7 using 68385-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HuH-7 cells using PFKL antibody (68385-1-Ig, Clone: 1H2G2 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HEK-293 using 68385-1-Ig

  1x10^6 HEK-293 cells were intracellularly stained with 0.8 ug PFKL Monoclonal antibody (68385-1-Ig, Clone:1H2G2) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1)(red), or 0.8 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: LNCaP cells, HeLa cells, HEK-293 cells, Jurkat cells, K-562 cells, pig brain tissue, rabbit brain tissue, rat brain tissue, mouse brain tissue
• Positive IHC detected in: human lung cancer tissue, human liver tissue, human ovary tumor tissue, mouse liver tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HuH-7 cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below

Product Information

68385-1-Ig targets PFKL in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with Human, Mouse, Rat, Rabbit, Pig samples.

• Tested Reactivity: Human, Mouse, Rat, Rabbit, Pig
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: PFKL fusion protein Ag8250
• Full Name: phosphofructokinase, liver
• Calculated Molecular Weight: 85 kDa
• Observed Molecular Weight: 85 kDa
• GenBank Accession Number: BC008964
• Gene Symbol: PFKL
• Gene ID (NCBI): 5211
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

PFKL (6-phosphofructokinase, liver type) is also named as PFK-B and belongs to the phosphofructokinase family. The PFKL gene encodes the liver isoform of phosphofructokinase (PFK) (ATP: D-fructose-6-phosphate-1-phosphotransferase). PFK catalyzes the irreversible conversion of fructose-6-phosphate to fructose-1,6-bisphosphate and is a key regulatory enzyme in glycolysis. In human beings, PFK exists as a system of three subunits: these are the muscle(PFKM), liver(PFKL), and platelet(PFKP) PFKs (PMID: 2139864). It has 2 isoforms produced by alternative splicing.

Publications

Species	Application	Title
human	WB	BMC Cardiovasc Disord; Identification of hub glycolysis-related genes in acute myocardial infarction and their correlation with immune infiltration using bioinformatics analysis; Authors - Xiaoqing Zhang
mouse	WB	Metabolism; Protein kinase N1 deficiency results in upregulation of cerebral energy metabolism and is highly protective in in vivo and in vitro stroke models; Authors - Stephanie Zur Nedden