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Anti-PCK2 Antibody Picoband™ (monoclonal, 3F7)

Figure 1. Western blot analysis of PCK2 using anti-PCK2 antibody (M04772-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEPG2 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human HEK293 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: monkey COS-7 whole cell lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse Neuro-2a whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-PCK2 antigen affinity purified monoclonal antibody (Catalog # M04772-2) at 0.25 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for PCK2 at approximately 71KD. The expected band size forPCK2 is at 71KD.
Figure 2. IHC analysis of PCK2 using anti-PCK2 antibody (M04772-2).
PCK2 was detected in paraffin-embedded section of human prostate cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-PCK2 Antibody (M04772-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 3. IHC analysis of PCK2 using anti-PCK2 antibody (M04772-2).
PCK2 was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-PCK2 Antibody (M04772-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 4. IHC analysis of PCK2 using anti-PCK2 antibody (M04772-2).
PCK2 was detected in paraffin-embedded section of human retcal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-PCK2 Antibody (M04772-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 5. IF analysis of PCK2 using anti-PCK2 antibody (M04772-2).
PCK2 was detected in immunocytochemical section of HEPG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-PCK2 Antibody (M04772-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 6. Flow Cytometry analysis of MCF-7 cells using anti-PCK2 antibody (M04772-2).
Overlay histogram showing MCF-7 cells stained with M04772-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PCK2 Antibody (M04772-2, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	Q16822
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse, Monkey
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-PCK2 Antibody Picoband™ (monoclonal, 3F7) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.
Reconstitution	Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Additional Information
Gene ID	5106
Other Names	Phosphoenolpyruvate carboxykinase [GTP], mitochondrial, PEPCK-M, 4.1.1.32, Phosphoenolpyruvate carboxykinase 2, mitochondrial, mtPCK2, PCK2 (HGNC:8725), PEPCK2
Calculated MW	70699 Da
Application Details	Western blot, 0.1-0.25 µg/ml, Human, Mouse, Rat, Monkey Immunohistochemistry (Paraffin-embedded Section), 2-5 µg/ml, Human Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Source	Eukaryota
Contents	Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clone Names	Clone: 3F7
Immunogen	E.coli-derived human PCK2 recombinant protein (Position: M1-M640).
Purification	Immunogen affinity purified.
Storage	Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	PCK2 (HGNC:8725)
Synonyms	PEPCK2
Function	Mitochondrial phosphoenolpyruvate carboxykinase that catalyzes the conversion of oxaloacetate (OAA) to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle (PubMed:28955899). Can play an active role in glyceroneogenesis and gluconeogenesis (PubMed:28955899). Also acts as a serine/threonine- protein kinase: phosphorylates and activates ACSL4, thereby promoting ferroptosis (PubMed:38720107).
Cellular Location	Mitochondrion
Tissue Location	Widely expressed..

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Phosphoenolpyruvate carboxykinase 2, mitochondrial (PCK2, PEPCK-M), is an isozyme of phosphoenolpyruvate carboxykinase (PCK, PEPCK) that in humans is encoded by the PCK2 gene. It is mapped to 14q11.2-q12. This gene encodes a mitochondrial enzyme that catalyzes the conversion of oxaloacetate to phosphoenolpyruvate in the presence of guanosine triphosphate (GTP). A cytosolic form of this protein is encoded by a different gene and is the key enzyme of gluconeogenesis in the liver. Alternatively spliced transcript variants have been described.

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¥ 3,367.00

Cat# ABO14980

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Anti-PCK2 Antibody Picoband™ (monoclonal, 3F7)

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