Apolipoprotein E Recombinant Rabbit Monoclonal Antibody [SC0536] - BSA and Azide free
Catalog# HA750206
Apolipoprotein E Recombinant Rabbit Monoclonal Antibody [SC0536] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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Human
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Rat
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ET1610-22
含抗保成分
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unconjugated
Overview
Product Name
Apolipoprotein E Recombinant Rabbit Monoclonal Antibody [SC0536] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within C-terminal human Apolipoprotein E.
Species Reactivity
Human, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP
Molecular Weight
Predicted band size: 36 kDa
Positive Control
Human plasma tissue lysates, human tonsil tissue, human liver tissue, human kidney tissue, human placenta tissue, human liver tissue, rat liver, HepG2.
Conjugation
unconjugated
Clone Number
SC0536
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:5,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:1,000
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IP
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Use at an assay dependent concentration.
Target
Function
Apolipoprotein-E (apoE) is a protein component of plasma lipoproteins that mediates the binding, internalization and catabolism of lipoprotein particles. It can serve as a ligand for several lipoprotein receptors, including the LDL (ApoB/E) receptor and the hepatic apoE (chylomicron remnant) receptor. apoE is produced in most organs and occurs in all plasma lipoprotein fractions, constituting 10-20% of VLDL (very low density lipoprotein) and 1-2% of HDL (high density lipoprotein). Three major isoforms of apoE have been described in human (E2, E3 and E4) which differ by only one or two amino acids. Estrogen receptor has been shown to upregulate apoE gene expression via the ERa-mediated pathway, indicating a potential role for apoE in atherosclerosis. This is consistent with studies in mice in which plasma apoE levels were raised, thereby protecting the mice from diet-induced atherosclerosis. apoE has also been shown to be a potent inhibitor of proliferation and thus may play a role in angiogenesis, tumor cell growth and metastasis.
Background References
1. Yang LG et al. Apolipoprotein E in lipid metabolism and neurodegenerative disease. Trends Endocrinol Metab. 2023 Aug
2. Saito T et al. Apolipoprotein E-related glomerular disorders. Kidney Int. 2020 Feb
Sequence Similarity
Belongs to the apolipoprotein A1/A4/E family.
Tissue Specificity
Produced by several tissues and cell types and mainly found associated with lipid particles in the plasma, the interstitial fluid and lymph. Mainly synthesized by liver hepatocytes. Significant quantities are also produced in brain, mainly by astrocytes and glial cells in the cerebral cortex, but also by neurons in frontal cortex and hippocampus. It is also expressed by cells of the peripheral nervous system. Also expressed by adrenal gland, testis, ovary, skin, kidney, spleen and adipose tissue and macrophages in various tissues.
Post-translational Modification
APOE exists as multiple glycosylated and sialylated glycoforms within cells and in plasma. The extent of glycosylation and sialylation are tissue and context specific. Plasma APOE undergoes desialylation and is less glycosylated and sialylated than the cellular form. Glycosylation is not required for proper expression and secretion. O-glycosylated with core 1 or possibly core 8 glycans. Thr-307 and Ser-314 are minor glycosylation sites compared to Ser-308.; Glycated in plasma VLDL of normal subjects, and of hyperglycemic diabetic patients at a higher level (2-3 fold).; Phosphorylated by FAM20C in the extracellular medium.; Undergoes C-terminal proteolytic processing in neurons. C-terminally truncated APOE has a tendency to form neurotoxic intracellular neurofibrillary tangle-like inclusions in neurons.
Subcellular Location
Secreted, extracellular space, extracellular matrix, Extracellular vesicle, Endosome, multivesicular body.
Synonyms
AD2 antibody
Apo-E antibody
APOE antibody
APOE_HUMAN antibody
APOEA antibody
Apolipoprotein E antibody
Apolipoprotein E3 antibody
ApolipoproteinE antibody
Apoprotein antibody
LDLCQ5 antibody
ExpandImages
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Western blot analysis of Apolipoprotein E on human plasma tissue lysates with Rabbit anti-Apolipoprotein E antibody (HA750206) at 1/1,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 36 kDa
Observed band size: 33 kDa
Exposure time: 6 seconds;
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 (HA750206) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Apolipoprotein E antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Apolipoprotein E antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Apolipoprotein E antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Apolipoprotein E antibody (HA750206) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Western blot analysis of Apolipoprotein E on different lysates with Rabbit anti-Apolipoprotein E antibody (HA750206) at 1/2,000 dilution.
Lane 1: HepG2 cell lysate
Lane 2: Human liver tissue lysate
Lane 3: Human plasma tissue lysate
Lysates/proteins at 20 µg/Lane1 and 40ug/Lane2-3.
Predicted band size: 35 kDa
Observed band size: 35 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 (HA750206) 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. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Apolipoprotein E antibody (HA750206) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Apolipoprotein E antibody (HA750206) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750206) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of HepG2 cells labeling Apolipoprotein E with Rabbit anti-Apolipoprotein E antibody (HA750206) 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-Apolipoprotein E antibody (HA750206) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HepG2 cells labeling Apolipoprotein E.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750206, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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