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PCK2 Recombinant Rabbit Monoclonal Antibody [JB52-39]

RMB: 900 特惠 1500

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Catalog# ET7107-29

PCK2 Recombinant Rabbit Monoclonal Antibody [JB52-39]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

PCK2 Recombinant Rabbit Monoclonal Antibody [JB52-39]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Recombinant protein within C-terminal Human PCK2 .

种属反应性

Human, Mouse, Rat

验证应用

WB, IF-Cell, IF-Tissue, IHC-P, FC

分子量

Predicted band size: 71 kDa

阳性对照

Mouse colon tissue lysate, MCF-7 cell lysate, human kidney tissue lysate, MCF7, Neuro-2a, mouse colon tissue, human kidney tissue, human liver tissue, rat kidney tissue, human colon cancer tissue.

偶联

unconjugated

克隆号

JB52-39

RRID

AB_3070742

产品特性

形态

Liquid

浓度

存放说明

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.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

靶点

功能

As a PCK, PCK2 catalyzes the GTP-driven conversion of OAA to PEP as a rate-limiting step in gluconeogenesis. This conversion step serves as a bridge between glycolytic and TCA cycle intermediates in the mitochondria. In pancreatic β-cells, PCK2 regulates glucose-stimulated insulin secretion by recycling GTP generated from the succinyl-CoA synthase reaction. This drives the TCA cycle, converting PEP to pyruvate to acetyl-CoA for the citrate synthase reaction.[9] Since nearly all of the glycolytic reactions upstream of PEP and downstream of glucose-6-phosphate (G6P) are reversible, PCK2-mediated synthesis of PEP could fuel multiple biosynthetic processes, such as serine synthesis, glycerol synthesis, and nucleotide synthesis. Notably, PCK2 preferentially converts OAA derived from lactate and, thus, can promote biosynthesis even under low-glucose conditions. As a result, PCK2 activity contributes to cell growth and survival during stress. While PCK1 is mainly expressed in the liver and kidney, PCK2 is ubiquitously expressed in various cell types, including leukocytes and neurons, as well as in non-gluconeogenic tissues, including pancreas, brain, heart. Moreover, while PCK1 expression is regulated by hormones or nutrients involved in gluconeogenesis, PCK2 is constitutively expressed. These differences indicate that PCK2 may also perform non-gluconeogenic functions.

背景文献

1. Ren Z et al. Enhancement of porcine intramuscular fat content by overexpression of the cytosolic form of phosphoenolpyruvate carboxykinase in skeletal muscle. Sci Rep 7:43746 (2017).

2. Méndez-Lucas A et al. Mitochondrial PEPCK is a Pro-Survival, ER-Stress Response Gene Involved in Tumor Cell Adaptation to Nutrient Availability. J Biol Chem 289(32):22090-102 (2014).

序列相似性

Belongs to the phosphoenolpyruvate carboxykinase [GTP] family.

亚细胞定位

Mitochondrion.

UNIPROT

SwissProt: Q16822 Human

SwissProt: Q8BH04 Mouse

Entrez Gene: 361042 Rat

别名

EC 4.1.1.32 antibody

GTP mitochondrial precursor antibody

HGNC:8725 antibody

mitochondrial antibody

Mitochondrial phosphoenolpyruvate carboxykinase 2 antibody

OTTHUMP00000164700 antibody

PCK2 antibody

PCKGM_HUMAN antibody

PE antibody

PEP carboxykinase antibody

展开

图片

  • Western blot analysis of PCK2 on different lysates with Rabbit anti-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 4: C6 cell lysate (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 6: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 71 kDa<br />Observed band size: 67 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PCK2 on different lysates with Rabbit anti-PCK2 antibody (ET7107-29) at 1/2,000 dilution.

    Lane 1: MCF7 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: Neuro-2a cell lysate (20 µg/Lane)
    Lane 4: C6 cell lysate (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (40 µg/Lane)
    Lane 6: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 71 kDa
    Observed band size: 67 kDa

    Exposure time: 20 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 (ET7107-29) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PCK2 on different lysates with Rabbit anti-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-PCK2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 71 kDa<br />Observed band size: 67 kDa<br /><br />Exposure time: 21 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PCK2 on different lysates with Rabbit anti-PCK2 antibody (ET7107-29) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-PCK2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 71 kDa
    Observed band size: 67 kDa

    Exposure time: 21 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 (ET7107-29) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of MCF7 cells labeling PCK2 with Rabbit anti-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/100 dilution.<br /><br />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-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of MCF7 cells labeling PCK2 with Rabbit anti-PCK2 antibody (ET7107-29) 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-PCK2 antibody (ET7107-29) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of Neuro-2a cells labeling PCK2 with Rabbit anti-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/100 dilution.<br /><br />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-PCK2 antibody (<a href="/products/ET7107-29" style="font-weight: bold;text-decoration: underline;">ET7107-29</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of Neuro-2a cells labeling PCK2 with Rabbit anti-PCK2 antibody (ET7107-29) 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-PCK2 antibody (ET7107-29) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-PCK2 antibody. Counter stained with hematoxylin.

请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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