Invitrogen

ApoA1 Polyclonal Antibody

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Questions & Answers

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ApoA1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis using Anti-ApoA1 Polyclonal Antibody (Product # PA5-29557), shows higher expression of ApoA1 in Hep G2 cell line as compared to SH-SY5Y. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ApoA1 Polyclonal Antibody (Product # PA5-29557) at 5 µg in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Blue). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image of untreated HepG2 cells showing staining for APOA1 that is enhanced upon PTI treatment (Panel h). Panel l represents SH-SY 5Y cells showing no staining for APOA1 upon PTI treatment. The images were captured at 60X magnification. APOA1 is secretory protein showing cytoplasmic and Golgi apparatus expression as seen in panel d and h respectively upon PTI treatment.

ApoA1 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry-Immunofluorescence analysis of ApoA1 was performed in HepG2 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: ApoA1 Polyclonal Antibody (Product # PA5-29557) diluted at 1:500. Blue: Hoechst 33342 staining.

ApoA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of ApoA1 was performed in paraffin-embedded human ovarian cancer tissue using ApoA1 Polyclonal Antibody (Product # PA5-29557) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

ApoA1 Antibody in Western Blot (WB) — Western blot was performed using Anti-ApoA1 Polyclonal Antibody (Product # PA5-29557) and a 25kDa band corresponding to ApoA1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with FLI, 10 uM, 4 Hrs. (Lane 2), Hep G2 treated with PTI, 1X, 4Hrs. (Lane 3), SH-SY5Y (Lane 4), SH-SY5Y treated with FLI, 10 uM, 4 Hrs (Lane 5), SH-SY5Y treated with PTI, 1X, 4Hrs. (Lane 6), Empty lane (Lane 7) and Human Plasma (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Expression of APOA1 was found to be higher in Hep G2 cell line as compared to SH-SY 5Y.

ApoA1 Antibody in Western Blot (WB) — Western blot analysis of ApoA1 was performed by separating 50 µg of mouse tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a ApoA1 Polyclonal Antibody (Product # PA5-29557) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

ApoA1 Antibody in Western Blot (WB) — Western blot analysis of ApoA1 was performed by separating 50 µg of rat tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a ApoA1 Polyclonal Antibody (Product # PA5-29557) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

ApoA1 Antibody in Western Blot (WB) — Western blot analysis of ApoA1 was performed by separating 30 µg of human plasma by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a ApoA1 Polyclonal Antibody (Product # PA5-29557) at a dilution of 1:10000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

ApoA1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines tested owing to their inherent genetic constitution. Expression of APOA1 was found to be higher in Hep G2 cell line as compared to SH-SY 5Y using Anti-ApoA1 Polyclonal Antibody (Product # PA5-29557) in Western Blot. {RE}

ApoA1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Immunofluorescence analysis using Anti-ApoA1 Polyclonal Antibody (Product # PA5-29557) shows expression of APOA1 higher in Hep G2 cell line as compared to SH-SY 5Y. {RE}

Product Details

PA5-29557

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:20,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant fragment corresponding to a region within amino acids 1 and 267 of Human Apolipoprotein A1 View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.43 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 1% BSA, 20% glycerol
Contains	0.025% ProClin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2547033

Product Specific Information

Recommended positive controls: human plasma, mouse plasma, rat plasma.

Predicted reactivity: Pig (80%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

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

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

UniProt ID: (Human) P02647, (Mouse) Q00623, (Rat) P04639

Entrez Gene ID: (Human) 335, (Mouse) 11806, (Rat) 25081

Molecular Function: apolipoprotein

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

Process(es)

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