Anti-Apolipoprotein A I/APOA1 Antibody Picoband™

Product Name

Anti-Apolipoprotein A I/APOA1 Antibody Picoband™

View all Apolipoprotein A-I/ApoA1 Antibodies

SKU/Catalog Number

PB9916

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Apolipoprotein A I/APOA1 Antibody Picoband™ catalog # PB9916. Tested in Flow Cytometry, IHC, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Apolipoprotein A I/APOA1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9916)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E. coli-derived human APOA1 recombinant protein (Position: D25-Q267). Human APOA1 shares 64% and 61.7% amino acid (aa) sequence identity with mouse and rat APOA1, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9916 is reactive to APOA1 in Human

Applications

PB9916 is guaranteed for Flow Cytometry, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

26 kDa

Calculated molecular weight

30.778kDa

Background of Apolipoprotein A-I/ApoA1

Apolipoprotein A-1, also known as APOA1, is a human protein with a specific role in lipid metabolism. It binds to lipopolysaccharide or endotoxin, and has a major role in the anti-endotoxin function of HDL. The gene is mapped to 11q23. And it is a single polypeptide chain with 243 amino acid residues of known primary amino acid sequence. The ApoA-I protein promotes cholesterol efflux from tissues to the liver for excretion. It is a cofactor for lecithin cholesterolacyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. ApoA-I is also isolated as a prostacyclin (PGI2) stabilizing factor, and thus may have an anticlotting effect. Defects in the gene encoding it are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis. Additionally, ApoA-I overexpression promotes macrophage-specific reverse cholesterol transport.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
ELISA , 0.1-0.5μg/ml, Human, -
Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of APOA1 using anti-APOA1 antibody (PB9916).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-APOA1 antigen affinity purified polyclonal antibody (Catalog # PB9916) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for APOA1 at approximately 26 kDa. The expected band size for APOA1 is at 31 kDa.

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Figure 2. IHC analysis of APOA1 using anti-APOA1 antibody (PB9916).
APOA1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-APOA1 Antibody (PB9916) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of APOA1 using anti-APOA1 antibody (PB9916).
APOA1 was detected in a paraffin-embedded section of human renal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-APOA1 Antibody (PB9916) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of CACO-2 cells using anti-APOA1 antibody (PB9916).
Overlay histogram showing CACO-2 cells stained with PB9916 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-APOA1 Antibody (PB9916,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 5. Flow Cytometry analysis of HepG2 cells using anti-APOA1 antibody (PB9916).
Overlay histogram showing HepG2 cells stained with PB9916 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-APOA1 Antibody (PB9916,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For APOA1 (Source: Uniprot.org, NCBI)

Alternative Names

Alp-1; APOA1; Apo-AI; apoA-I; Apolipoprotein A1; Apolipoprotein AI; Apolipoprotein A-I; Brp-14; Ltw-1; Lvtw-1; MGC117399; Sep-1; Sep-2 APOA1 HPALP2, apo(a) apolipoprotein A1 apolipoprotein A-I|apo-AI|epididymis secretory sperm binding protein

For more info on APOA1, check out the APOA1 Infographic

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In this infographic, you will see the following information for APOA1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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