Anti-P Glycoprotein/ABCB1 Antibody Picoband™

Product Name

Anti-P Glycoprotein/ABCB1 Antibody Picoband™

View all MDR1/ABCB1 Antibodies

SKU/Catalog Number

A00049-4

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-P Glycoprotein/ABCB1 Antibody Picoband™ catalog # A00049-4. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-P Glycoprotein/ABCB1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00049-4)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human P Glycoprotein/ABCB1, which shares 78.9% amino acid (aa) sequence identity with mouse ABCB1.

*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

A00049-4 is reactive to ABCB1 in Human

Applications

A00049-4 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

141 kDa

Calculated molecular weight

141.479kDa

Background of MDR1/ABCB1

P-GP, also called ABCB1 or PGY1, is a glycoprotein that in humans is encoded by the ABCB1 gene. It is mapped to 7q21.12. P-GP is a well-characterized ABC-transporter (which transports a wide variety of substrates across extra- and intracellular membranes) of the MDR/TAP subfamily. It is an important protein of the cell membrane that pumps many foreign substances out of cells. More formally, it is an ATP-dependent drug efflux pump with broad substrate specificity. P-GP is an ATP-dependent drug efflux pump forxenobiotic compounds with broad substrate specificity. It is responsible for decreased drug accumulation in multidrug-resistant cells and often mediates the development of resistance to anticancer drugs. This protein also functions as a transporter in the blood–brain barrier.

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

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/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.

Western blot, 0.25-0.5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of P Glycoprotein/ABCB1 using anti-P Glycoprotein/ABCB1 antibody (A00049-4).
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 HepG2 whole cell lysates,
Lane 2: human Caco-2 whole cell 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-P Glycoprotein/ABCB1 antigen affinity purified polyclonal antibody (Catalog # A00049-4) 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 P Glycoprotein/ABCB1 at approximately 141 kDa. The expected band size for P Glycoprotein/ABCB1 is at 141 kDa.

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Figure 2. IF analysis of P Glycoprotein/ABCB1 using anti-P Glycoprotein/ABCB1 antibody (A00049-4).
P Glycoprotein/ABCB1 was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-P Glycoprotein/ABCB1 Antibody (A00049-4) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of 293T cells using anti-P Glycoprotein/ABCB1 antibody (A00049-4).
Overlay histogram showing 293T cells stained with A00049-4 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-P Glycoprotein/ABCB1 Antibody (A00049-4, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For ABCB1 (Source: Uniprot.org, NCBI)

Alternative Names

ABC20; ABCB1; ABCB1B; ATP-binding cassette sub-family B member 1; ATP-binding cassette, sub-family B (MDR/TAP), member 1; CD243 antigen; CD243; CLCS; colchicin sensitivity; doxorubicin resistance; EC 3.6.3; EC 3.6.3.44; GP170; IBD13; MDR1; MDR1MGC163296; multidrug resistance protein 1; P-glycoprotein 1; P-gp; PGY1; PGY1P-GP ABCB1 ABC20, CD243, CLCS, GP170, MDR1, P-GP, PGY1, p-170 ATP binding cassette subfamily B member 1 ATP-dependent translocase ABCB1|ATP-binding cassette, sub-family B (MDR/TAP), member 1|P-glycoprotein 1|colchicin sensitivity|doxorubicin resistance|multidrug resistance protein 1|phospholipid transporter ABCB1

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