Invitrogen

ApoA1 Monoclonal Antibody (532)

查看其他48个ApoA1抗体

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ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

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ApoA1 Antibody (MIA1405) in IHC (P)

Immunohistochemistry analysis of Apolipoprotein A-1 showing staining in the cytoplasm of paraffin-embedded human small intestine tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min a... View More

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ApoA1 Antibody in Western Blot (WB)

ApoA1 Antibody in Western Blot (WB)

ApoA1 Antibody in Western Blot (WB)

ApoA1 Antibody in ELISA (ELISA)

ApoA1 Antibody in ELISA (ELISA)

ApoA1 Antibody in ELISA (ELISA)

ApoA1 Antibody in ELISA (ELISA)

Product Image Gallery

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Apolipoprotein A-1 showing staining in the cytoplasm of paraffin-embedded human small intestine tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Apolipoprotein A-1 Mouse Monoclonal Antibody (Product # MIA1405) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Apolipoprotein A-1 showing staining in the cytoplasm of paraffin-embedded mouse small intestine tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Apolipoprotein A-1 Mouse Monoclonal Antibody (Product # MIA1405) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Apolipoprotein A-1 showing staining in the cytoplasm of paraffin-embedded human liver tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Apolipoprotein A-1 Mouse Monoclonal Antibody (Product # MIA1405) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on human liver tissue sections. To expose Apo A-1, tissues were microwaved for 10 minutes in 10mM sodium citrate, pH 6.0. Following antigen retrieval, endogenous peroxidases were blocked in a 3% hydrogen peroxide-methanol solution for 15 minutes, and then tissues were blocked in 3% BSA in PBS for 30 minutes at room temperature. Tissues were probed with an Apo A-1 monoclonal antibody (Product # MIA1405) at a dilution of 1:100 overnight at 4C, washed extensively in PBST, and detection was performed with an HRP-conjugated anti-mouse IgG secondary antibody followed by DAB substrate. Tissues were counterstained with hematoxylin.

ApoA1 Antibody in Western Blot (WB) — Fig 1: Western blot analysis was performed on whole cell and tissue extracts (30 µg lysate) of Hep G2 (Lane 1) and Mouse Liver (Lane 2). Fig 2: Likewise, Western blot analysis was performed on samples of 3 µL serum (Lane 1), 5 µL serum (Lane 2), 5 µL plasma (Lane 3), 1 µL plasma (Lane 4), all of which were obtained from a 50 year old male Alzheimer's patient, and 10 µL of conditioned media from HepG2 cell line (Lane 5). The blots were probed with Anti-Apolipoprotein A-1 Mouse Monoclonal Antibody (Product # MIA1405, 1:500-1:2000 dilution) and detected by chemiluminescence Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 28 kDa band corresponding to Apolipoprotein A-1 was observed across the cell line, tissue and samples tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

ApoA1 Antibody in Western Blot (WB) — Western blot analysis of Apolipoprotein A-1 was performed by loading 10 µL of serum depleted of highly abundant proteins using the Top 2 Abundant Protein Depletion Spin Columns (Product # 85162, left lane) or the Top 12 Abundant Protein Depletion Spin Columns (Product # 85164, middle lane) and concentrated using the PES Protein Concentrators, 10K MWCO (Product # 88513), 50 µg of HepG2 whole cell lysate (right lane), and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX ). Proteins were transferred to a Nitrocellulose Membrane (Product # 88014) using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. Apo A-1 was detected at ~25 kD using an Apolipoprotein A-1 monoclonal antibody (Product # MIA1405) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG Fc-specific secondary antibody (Product # 31439) at a dilution of 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

ApoA1 Antibody in Western Blot (WB) — Western blot analysis of Apolipoprotein A-1 was performed by loading the indicated amounts of a recombinant human Apo A-1-hIgG1Fc fusion protein (Product # 10686-H02H-50) or a recombinant mouse Apo A-1-hIgG1Fc fusion protein (Product # 50918-M02H-50), and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX ). Proteins were transferred to a Nitrocellulose Membrane (Product # 88014) using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. Apo A-1 was detected at ~50 kD using an Apolipoprotein A-1 monoclonal antibody (Product # MIA1405) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG Fc-specific secondary antibody (Product # 31439) at a dilution of 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

ApoA1 Antibody in ELISA (ELISA) — Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL of diluted normal human serum or diluted serum from a patient with dyslipidemia (B) were incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MIA1405) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated Rabbit anti-Mouse IgG Superclonal secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

ApoA1 Antibody in ELISA (ELISA) — Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 rabbit monoclonal antibody (Product # 701239) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL of diluted normal human serum or diluted serum from a patient with dyslipidemia (B) were incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 mouse monoclonal antibody (Product # MIA1405) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated rabbit anti-mouse IgG Superclonal™ secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

ApoA1 Antibody in ELISA (ELISA) — Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000 ng/mL and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MIA1405) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of an HRP-conjugated Rabbit anti-Mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

ApoA1 Antibody in ELISA (ELISA) — Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 rabbit monoclonal antibody (Product # 701239) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000 ng/mL and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 mouse monoclonal antibody (Product # MIA1405) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of an HRP-conjugated rabbit anti-mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

ApoA1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target between normal and disease condition. Higher levels of Apolipoprotein-A1 expression were observed in serum from a dyslipidemic patient compared to normal serum using Apolipoprotein-A1 Monoclonal Antibody (Product # MIA1405) in ELISA. {RE}

ApoA1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target between normal and disease condition. Higher levels of Apolipoprotein-A1 expression were observed in serum from a dyslipidemic patient compared to normal serum using Apolipoprotein-A1 Monoclonal Antibody (Product # MIA1405) in ELISA. {RE}

产品信息

MIA1405

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:500-1:2,000

-

免疫组化（石蜡） (IHC (P))

1:20-1:100

-

酶联免疫吸附实验 (ELISA)

1 µg/mL

-

免疫沉淀 (IP)

Assay-dependent

-

放射免疫测定 (RIA)

Assay-dependent

-

产品规格

种属反应

Human, Mouse

宿主/亚型

Mouse / IgG1

分类

Monoclonal

类型

Antibody

克隆号

532

抗原

Purified human plasma Apo A-1

偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

5 mg/mL

纯化类型

Protein A

保存液

PBS, pH 7.4

内含物

0.09% sodium azide

保存条件

Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C

运输条件

Ambient (domestic); Wet ice (international)

AB_11152905

产品详细信息

By sandwich ELISA, MIA1405 can be used as a detection antibody with Product # 710263 or # 701239 as a coating antibody, to generate a matched pair. Using these matched pairs, recombinant human Apo A-1, but not recombinant mouse Apo A-1, was detected. MIA1405 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MIA1405, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.

By Western blot, MIA1405 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MIA1405 is also recommended for detecting endogenous Apo A-1 by Western blot.

MIA1405 was formerly sold as a Seradyn product.

靶标信息

Apolipoprotein A-1 (apo A-1) in human plasma is a major protein component of high density lipoprotein (HDL). The molecular mass is 24 kDa. Apo A-1 is a cofactor for enzymes involved in the normal turnover of cholesterol and is produced in the liver. Plasma concentration is 270-380 mg/100 mL.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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生物信息学

蛋白别名: 1-Sep; 2-Sep; Apo a; apo A-1; APO A1; APO I; Apo-AI; ApoAI; Apolipoprotein; Apolipoprotein A-I; Apolipoprotein A-I(1-242); Apolipoprotein A1; Apolipoprotein AI; MGC102525; MGC117399; MGC124929; ProapoA-I; Proapolipoprotein A-I; Truncated apolipoprotein A-I; Udpgt; Ugt1; Ugt1a7

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基因别名: Alp-1; apo(a); apo-AI; Apoa-1; apoA-I; APOA1; Brp-14; Ltw-1; Lvtw-1; Sep-1; Sep-2; Sep2

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UniProt ID: (Human) P02647, (Mouse) Q00623

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Entrez Gene ID: (Human) 335, (Mouse) 11806

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beta-amyloid binding protein binding phospholipid binding phospholipid transporter activity high-density lipoprotein particle binding cholesterol binding cholesterol transporter activity enzyme binding phosphatidylcholine binding apolipoprotein receptor binding apolipoprotein A-I receptor binding identical protein binding chemorepellent activity lipase inhibitor activity phosphatidylcholine-sterol O-acyltransferase activator activity high-density lipoprotein particle receptor binding lipid transporter activity lipid binding lipoprotein particle binding

regulation of protein phosphorylation endothelial cell proliferation negative regulation of cytokine secretion involved in immune response lipid metabolic process phospholipid metabolic process phosphatidylcholine biosynthetic process cholesterol biosynthetic process transport lipid transport G-protein coupled receptor signaling pathway integrin-mediated signaling pathway protein localization steroid metabolic process cholesterol metabolic process glucocorticoid metabolic process negative regulation of tumor necrosis factor-mediated signaling pathway positive regulation of cholesterol esterification positive regulation of triglyceride catabolic process negative regulation of very-low-density lipoprotein particle remodeling phospholipid transport protein oxidation peptidyl-methionine modification triglyceride catabolic process lipid storage regulation of intestinal cholesterol absorption cholesterol transport adrenal gland development organ regeneration axon regeneration regulation of Cdc42 protein signal transduction cholesterol efflux phospholipid efflux negative regulation of heterotypic cell-cell adhesion very-low-density lipoprotein particle remodeling high-density lipoprotein particle remodeling high-density lipoprotein particle assembly positive regulation of Rho protein signal transduction lipoprotein metabolic process lipoprotein biosynthetic process cholesterol homeostasis blood vessel endothelial cell migration reverse cholesterol transport positive regulation of fatty acid biosynthetic process phosphatidylcholine metabolic process negative regulation of interleukin-1 beta secretion negative regulation of inflammatory response protein stabilization negative chemotaxis positive regulation of lipoprotein lipase activity vitamin transport positive regulation of hydrolase activity negative regulation of hydrolase activity positive regulation of transferase activity positive regulation of stress fiber assembly phospholipid homeostasis negative regulation of lipase activity negative regulation of cell adhesion molecule production negative regulation of response to cytokine stimulus triglyceride homeostasis cholesterol import positive regulation of substrate adhesion-dependent cell spreading retinoid metabolic process platelet degranulation receptor-mediated endocytosis transforming growth factor beta receptor signaling pathway response to nutrient peripheral nervous system axon regeneration neuron projection regeneration high-density lipoprotein particle clearance response to drug response to estrogen cellular protein metabolic process transmembrane transport ERK1 and ERK2 cascade