Western Blot (WB)
(Figure 1. Western blot analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).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 50ug of sample under reducing conditions.Lane 1: MCF-7 Whole Cell Lysate,Lane 2: SW620 Whole Cell Lysate,Lane 3: 22RV1 Whole Cell Lysate,Lane 4: SKOV Whole Cell Lysate.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-ABCB4 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ABCB4 at approximately 142KD. The expected band size for ABCB4 is at 142KD. )
Rabbit ABCB4 Polyclonal Antibody | anti-ABCB4 antibody
Anti-ABCB4 Antibody
Concentration: 0.1-0.5ug/ml
Tested Species: Hu, Ms, Rat
IHC (Paraffin-embedded Section):
Concentration: 0.5-1ug/ml
Tested Species: Hu
Antigen Retrieval: By Heat
IF/ICC:
Concentration: 2 ug/ml
Tested Species: Hu
Flow Cytometry:
1-3ug/1x106 cels
Tested Species: Hu
WB: The detection limit for ABCB4 is approximately 0.1ng/lane under reducing conditions.
Tested Species: In-house tested species with positive results.
By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20 mins is required for the staining of formalin/paraffin sections.
Other applications have not been tested.
Optimal dilutions should be determined by end users.
Western Blot (WB)
(Figure 1. Western blot analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).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 50ug of sample under reducing conditions.Lane 1: MCF-7 Whole Cell Lysate,Lane 2: SW620 Whole Cell Lysate,Lane 3: 22RV1 Whole Cell Lysate,Lane 4: SKOV Whole Cell Lysate.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-ABCB4 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ABCB4 at approximately 142KD. The expected band size for ABCB4 is at 142KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).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 50ug of sample under reducing conditions.Lane 1: MCF-7 Whole Cell Lysate,Lane 2: SW620 Whole Cell Lysate,Lane 3: 22RV1 Whole Cell Lysate,Lane 4: SKOV Whole Cell Lysate.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-ABCB4 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ABCB4 at approximately 142KD. The expected band size for ABCB4 is at 142KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Mouse Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Mouse Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Rat Liver Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Rat Liver Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Human Mammary Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of ABCB4 using anti-ABCB4 antibody (MBS177897).ABCB4 was detected in paraffin-embedded section of Human Mammary Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-ABCB4 Antibody (MBS177897) 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) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of A375 cells using anti-MDR3 antibody (MBS177897).Overlay histogram showing A375 cells stained with MBS177897 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MDR3 Antibody (MBS177897,1ug/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of A375 cells using anti-MDR3 antibody (MBS177897).Overlay histogram showing A375 cells stained with MBS177897 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MDR3 Antibody (MBS177897,1ug/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of A549 cells using anti-MDR3 antibody (MBS177897).Overlay histogram showing A549 cells stained with MBS177897 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MDR3 Antibody (MBS177897,1ug/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of A549 cells using anti-MDR3 antibody (MBS177897).Overlay histogram showing A549 cells stained with MBS177897 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MDR3 Antibody (MBS177897,1ug/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Background: Adenosine triphosphate-binding cassette subfamily B, member 4 (ABCB4), also called MDR3, is a protein that in humans is encoded by the ABCB4 gene. The membrane-associated protein encoded by this gene is a member of the superfamily of ATP-binding cassette (ABC) transporters. The ABCB4 gene contains 28 exons and 27 of these contain coding sequences for the two homologous halves of the protein that correlate with functional domains. ABCB4 gene mutations represent a genetic factor involved in this peculiar form of cholesterol gallstone disease in adults.
NCBI and Uniprot Product Information
NCBI Description
The membrane-associated protein encoded by this gene is a member of the superfamily of ATP-binding cassette (ABC) transporters. ABC proteins transport various molecules across extra- and intra-cellular membranes. ABC genes are divided into seven distinct subfamilies (ABC1, MDR/TAP, MRP, ALD, OABP, GCN20, White). This protein is a member of the MDR/TAP subfamily. Members of the MDR/TAP subfamily are involved in multidrug resistance as well as antigen presentation. This gene encodes a full transporter and member of the p-glycoprotein family of membrane proteins with phosphatidylcholine as its substrate. The function of this protein has not yet been determined; however, it may involve transport of phospholipids from liver hepatocytes into bile. Alternative splicing of this gene results in several products of undetermined function. [provided by RefSeq, Jul 2008]
Uniprot Description
ABCB4: a transporter that mediates ATP-dependent export of organic anions and drugs from the cytoplasm. Hydrolyzes ATP with low efficiency. Human ABCB4 is not capable of conferring drug resistance. Mediates the translocation of phosphatidylcholine across the canalicular membrane of the hepatocyte. Interacts with HAX1. Defects are the cause of gallstones, and progressive familial intrahepatic cholestasis 3, an autosomal recessive liver disorder that progresses to cirrhosis and liver failure before adulthood. Two alternatively spliced human isoforms have been described.
Protein type: Transporter, ABC family; EC 3.6.3.44; Hydrolase; Transporter; Membrane protein, integral; Membrane protein, multi-pass
Chromosomal Location of Human Ortholog: 7q21.1
Cellular Component: apical plasma membrane; clathrin-coated vesicle; cytoplasm; integral to plasma membrane; intercellular canaliculus; lipid raft; membrane; plasma membrane
Molecular Function: ATP binding; ATPase activity, coupled to transmembrane movement of substances; phosphatidylcholine transmembrane transporter activity; phospholipid transporter activity; protein binding; xenobiotic-transporting ATPase activity
Biological Process: antigen processing and presentation of endogenous peptide antigen via MHC class I via ER pathway, TAP-dependent; antigen processing and presentation of endogenous peptide antigen via MHC class Ib via ER pathway, TAP-dependent; antigen processing and presentation of exogenous protein antigen via MHC class Ib, TAP-dependent; bile acid secretion; lipid homeostasis; lipid metabolic process; multidrug transport; phospholipid translocation; positive regulation of antigen processing and presentation of peptide antigen via MHC class I; positive regulation of cholesterol transport; response to drug; transmembrane transport; transport; xenobiotic transport
Disease: Cholestasis, Intrahepatic, Of Pregnancy 3; Cholestasis, Progressive Familial Intrahepatic, 3; Gallbladder Disease 1
Research Articles on ABCB4
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Product Notes
The ABCB4 abcb4 (Catalog #AAA177897) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-ABCB4 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's ABCB4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC-P) Paraffin, Immunofluoresence/Immunocytochemistry (IF/ICC), Flow Cytometry (FC/FACS) . WB: Concentration: 0.1-0.5ug/ml Tested Species: Hu, Ms, Rat IHC (Paraffin-embedded Section): Concentration: 0.5-1ug/ml Tested Species: Hu Antigen Retrieval: By Heat IF/ICC: Concentration: 2 ug/ml Tested Species: Hu Flow Cytometry: 1-3ug/1x106 cels Tested Species: Hu WB: The detection limit for ABCB4 is approximately 0.1ng/lane under reducing conditions. Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20 mins is required for the staining of formalin/paraffin sections. Other applications have not been tested. Optimal dilutions should be determined by end users. . Researchers should empirically determine the suitability of the ABCB4 abcb4 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ABCB4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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