Invitrogen

ApoA1 Monoclonal Antibody (513)

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

1 Published Figure

3 References

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ApoA1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Apolipoprotein A1 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Apolipoprotein A1 (513) Mouse Monoclonal Antibody (Product # MIA1404) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

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 # MIA1404) 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 Western Blot (WB) — Fig 1: Western blot analysis was performed on tissue extract (30 µg lysate) of Mouse Liver (Lane 1). 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), 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 4). The blots were probed with Anti-Apolipoprotein A-1 Mouse Monoclonal Antibody (Product # MIA1404, 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, tissues 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 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 # MIA1404) 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 Western Blot, Immunohistochemistry (Frozen) (WB, IHC (F)) — Activated ABAC1/ApoA1 signaling is involved in the antidepressant action of c-MSST targeting the left V1 of CUMS mice (A and B) Heat map of the differentially expressed proteins analyzed by iTRAQ (Group A, Control mice; Group B, CUMS mice; Group C, Sham c-MSST mice; Group D, c-MSST mice), n = 3 mice per group. (C and D) PPI network of the differential proteins in the four groups. The red circles indicate the major/hub nodes of the PPI network. (E and F) Representative fluorescence images of the mouse V1. Immunofluorescent double labeling for ApoA1 (red)/NeuN (green) and ABCA1 (green)/NeuN (red). Nuclei were stained blue with DAPI. n = 4 mice per group. Scale bar, 20 mum. (G) Colocalization of the three neurocytemarkers with ApoA1 was quantified in the left V1 of control and CUMS mice. n = 8-9 mice per group. Data are expressed as the mean +-SEM. ***p < 0.001 versus Con-NeuN+/ApoA1+; ### p < 0.001 versus CUMS-NeuN+/ApoA1+ using ANOVA with Sidak correction. (H-J) Expression of ApoA1 and ABCA1 in the left V1 of control and CUMS mice. n = 6 mice per group. Data are expressed as mean +-SEM. *p < 0.05, **p < 0.01, ***p < 0.001 using ANOVA with Bonferroni correction. iTRAQ, isobaric tagging for relative and absolute quantification; PPI, protein-protein interaction; ApoA1, apolipoprotein A1; ABCA1, ATP-binding cassette, subfamily A, member one; NeuN, neuronal nuclei; DAPI, 4',6-diamidino-2-phenylindole; SPIO, superparamagnetic iron oxide; MF, magnetic field; c-MSST, combined magnetic

ApoA1 Antibody — Antibody specificity was demonstrated by detection of relative expression of the target protein across patient samples owing to their inherent genetic constitution. Increased expression of Apolipoprotein A1 in serum from patients suffering from dyslipidemia was detected in ELISA using anti-Apolipoprotein A1 monoclonal antibody (513) (Product # MIA1404). {RE}

Product Details

MIA1404

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:2,000	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:100	-
Immunohistochemistry (Frozen) (IHC (F))	-	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	2 µg/mL	-
ELISA (ELISA)	1 µg/mL	-
Immunoprecipitation (IP)	Assay-dependent	-
Radioimmune Assays (RIA)	Assay-dependent	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	513
Immunogen	Purified human plasma Apo A-1
Conjugate	Unconjugated
Form	Liquid
Concentration	5 mg/mL
Purification	Ion-exchange chromatography
Storage buffer	PBS, pH 7.4
Contains	0.09% sodium azide
Storage conditions	Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C
RRID	AB_11151929

Product Specific Information

By sandwich ELISA, MIA1404 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. MIA1404 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MIA1404, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.

By Western blot, MIA1404 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MIA1404 is also recommended for detecting endogenous Apo A-1 by Western blot.

MIA1404 was formerly sold as a Seradyn product.

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.

Product References

Selective activation of ABCA1/ApoA1 signaling in the V1 by magnetoelectric stimulation ameliorates depression via regulation of synaptic plasticity.

iScience

Lu Q,Wu F,Jiao J,Xue L,Song R,Shi Y,Kong Y,Sun J,Gu N,Han MH,Zhang Z

Published figure using ApoA1 monoclonal antibody (Product # MIA1404) in Immunohistochemistry (Frozen)

Fri May 20 00:00:00 EDT 2022

Selective activation of ABCA1/ApoA1 signaling in the V1 by magnetoelectric stimulation ameliorates depression via regulation of synaptic plasticity.

iScience

Lu Q,Wu F,Jiao J,Xue L,Song R,Shi Y,Kong Y,Sun J,Gu N,Han MH,Zhang Z

Published figure using ApoA1 monoclonal antibody (Product # MIA1404) in Immunohistochemistry (Frozen)

Fri May 20 00:00:00 EDT 2022

Apolipoprotein A-I and B distribution in the human cornea.

Investigative ophthalmology & visual science

Ashraf F,Cogan DG,Kruth HS

MIA1404 was used in immunohistochemistry to study the expression of apoliprotein A1 and B in human cornea

Wed Dec 01 00:00:00 EST 1993

Bioinformatics

Protein Aliases: 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

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

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

Molecular Function: apolipoprotein