PCB Antibody [K5L1]

Catalog No.: F3292

Application: Reactivity: Mouse, Rat, Human

Lane 1: HepG2, Lane 2: HepG2 (KO PCB), Lane 3: Mouse brain, Lane 4: Rat brain

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代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

使用情報

Dilution
WB1:1000 - 1:10000

Application
WB

Source
Rabbit Monoclonal Antibody

Reactivity
Mouse, Rat, Human

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
130 kDa 130 kDa
^*なぜ予測分子量と実際の分子量が異なるのか？下記の原因により、実際の分子量が予測と異なる：タンパク質の翻訳後修飾（リン酸化/糖鎖付加），スプライシングバリアント，イソフォーム，相対的な電荷，ポリマー。

Datasheet & SDS

生物学的記述

Specificity
PCB Antibody [K5L1] detects endogenous levels of total PCB protein.

Clone
K5L1

Synonym(s)
Pyruvic carboxylase; PCB; PC

Background
Pyruvate carboxylase (PC, PCB) is a biotin-dependent mitochondrial enzyme that catalyzes the ATP- and CO₂-dependent carboxylation of pyruvate to oxaloacetate, functioning as the primary anaplerotic reaction to replenish TCA cycle intermediates and as the committed step of gluconeogenesis in the liver and kidney. PC forms a 500–550 kDa tetramer with tetrahedral symmetry, where each ~125 kDa subunit contains three domains: a biotin carboxylase (BC) domain for ATP/bicarbonate-dependent carboxybiotin formation, a carboxyl transferase (CT) domain with a Zn²⁺-stabilized pyruvate enolate site (key residues Asp549, Thr882, Lys718), and a biotin carboxyl carrier protein (BCCP) domain with a swinging biotinylated lysine that shuttles CO₂ between active sites approximately 55 Å apart. An allosteric acetyl-CoA binding domain induces domain closure, increasing enzyme Vmax up to 1000-fold through tetramer reorganization. PC operates via a two-step ping-pong mechanism: (1) the BC domain carboxylates biotin (HCO₃⁻ + MgATP → carboxybiotin), and (2) BCCP delivers the carboxyl to the CT domain, where carboxybiotin donates CO₂ to the pyruvate enolate, yielding oxaloacetate. Deficiency of PC leads to type A/B pyruvate carboxylase deficiency (characterized by lactic acidosis, hyperammonemia, and Leigh-like encephalopathy) due to TCA cycle impairment, while hepatic PC knockout protects against diet-induced hyperglycemia.

Background

Pyruvate carboxylase (PC, PCB) is a biotin-dependent mitochondrial enzyme that catalyzes the ATP- and CO₂-dependent carboxylation of pyruvate to oxaloacetate, functioning as the primary anaplerotic reaction to replenish TCA cycle intermediates and as the committed step of gluconeogenesis in the liver and kidney. PC forms a 500–550 kDa tetramer with tetrahedral symmetry, where each ~125 kDa subunit contains three domains: a biotin carboxylase (BC) domain for ATP/bicarbonate-dependent carboxybiotin formation, a carboxyl transferase (CT) domain with a Zn²⁺-stabilized pyruvate enolate site (key residues Asp549, Thr882, Lys718), and a biotin carboxyl carrier protein (BCCP) domain with a swinging biotinylated lysine that shuttles CO₂ between active sites approximately 55 Å apart. An allosteric acetyl-CoA binding domain induces domain closure, increasing enzyme Vmax up to 1000-fold through tetramer reorganization. PC operates via a two-step ping-pong mechanism: (1) the BC domain carboxylates biotin (HCO₃⁻ + MgATP → carboxybiotin), and (2) BCCP delivers the carboxyl to the CT domain, where carboxybiotin donates CO₂ to the pyruvate enolate, yielding oxaloacetate. Deficiency of PC leads to type A/B pyruvate carboxylase deficiency (characterized by lactic acidosis, hyperammonemia, and Leigh-like encephalopathy) due to TCA cycle impairment, while hepatic PC knockout protects against diet-induced hyperglycemia.

References
[1] Valle M. Subcell Biochem. 2017;83:291-322. [2] Jitrapakdee S, et al. Biochem J. 2008 Aug 1;413(3):369-87.

PVDFメンブレン孔径参考表

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%