Human BCRP/ABCG2 peptide (ab100820)

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Description

Specifications

Our Abpromise guarantee covers the use of ab100820 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Form

    Liquid

  • Additional notes

    - First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
    - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
    - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
    - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
    - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    Information available upon request.

General Info

  • Alternative names

    • ABC transporter
    • ABC15
    • ABCG 2
    • ABCG2
    • ABCG2_HUMAN
    • ABCP
    • ATP binding cassette sub family G (WHITE) member 2
    • ATP binding cassette transporter G2
    • ATP-binding cassette sub-family G member 2
    • BCRP
    • BCRP1
    • BMDP
    • Breast cancer resistance protein
    • CD338
    • CDw338
    • CDw338 antigen
    • EST157481
    • GOUT1
    • MGC102821
    • Mitoxantrone resistance associated protein
    • Mitoxantrone resistance-associated protein
    • MRX
    • Multi drug resistance efflux transport ATP binding cassette sub family G (WHITE) member 2
    • MXR
    • MXR1
    • Placenta specific ATP binding cassette transporter
    • Placenta specific MDR protein
    • Placenta-specific ATP-binding cassette transporter
    • UAQTL1
    see all

  • Function

    Xenobiotic transporter that may play an important role in the exclusion of xenobiotics from the brain. May be involved in brain-to-blood efflux. Appears to play a major role in the multidrug resistance phenotype of several cancer cell lines. When overexpressed, the transfected cells become resistant to mitoxantrone, daunorubicin and doxorubicin, display diminished intracellular accumulation of daunorubicin, and manifest an ATP-dependent increase in the efflux of rhodamine 123.

  • Tissue specificity

    Highly expressed in placenta. Low expression in small intestine, liver and colon.

  • Sequence similarities

    Belongs to the ABC transporter superfamily. ABCG family. Eye pigment precursor importer (TC 3.A.1.204) subfamily.
    Contains 1 ABC transmembrane type-2 domain.
    Contains 1 ABC transporter domain.

  • Post-translational
    modifications

    Glycosylation-deficient ABCG2 is normally expressed and functional.

  • Cellular localization

    Cell membrane.
  • Information by UniProt

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab100820? Please let us know so that we can cite the reference in this datasheet.

ab100820 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Question

We probed human placenta samples (60 ug) for the efflux transporter BCRP using your BCRP/ABCG2 primary antibody (ab63907, lot no: GR42347) for the first time. Our positive controls were mouse kidney lysates and human embryonic kidney (HEK) cells that overexpress the human BCRP. As a negative control we used the HEK cells that were overexpressing an empty vector. Additionally we added the same primary anitbody and the competing peptide (ab100820, lot no: GR53148) to another blot with the identical samples to be sure that the antibody is specific for BCRP. The positive controls (mouse kidney) did not show up, EV cells showed BCRP expression, and the competing peptide did not eliminate the bands. Below are some of the experimental conditions Blocking conditions: 5% milk in PBS/0.5% Tween, 1 hr Detection method: Chemiluminescence 20 minutes Electrophoresis: 160 V, 120 mA, 25 W 1.5 hours in 1x NuPage MOPS SDS Running Buffer (Life Technologies) Transfer: 7 minute dry transfer (Life Technologies) Primary ab: 1:1000 in 2% milk PBS/T overnight 4C and Primary + competing peptide (1:1000) Wash: 3x10 min PBS/T Secondary ab: 1:5000 in 2% milk PBS/T 1 hr RT Wash: 3x10 min PBS/T See powerpoint slide for images of blots. Thank you for your time.

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Answer

Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab63907 and ab100820. I would also appreciate if you can confirm some further details to help us better understand the problem with the antibody.

We haven't tested this antibody using mouse samples (only human and pig), so we are unsure how or if this antibody will work with mouse samples. We predict that the antibody may bind due to high sequence homology (>90%), but these are the first testing data we've seen so it may be that the antibody is not able to bind the mouse protein.

How were the samples prepared? For multiplass membrane proteins, we would recommend using RIPA buffer and heating to 70C for 5 - 10 mins rather than boiling since boiling can cause aggregates.

What percentage gel are you using? Also for the transfer, you may want to choose a wet transfer for these types of proteins. Make sure the transfer buffer has 0.1% SDS in it since this protein is hydrophobic and may have trouble entering the gel. This may explain why you are seeing such "wispy" bands in the blot.

What MW is that main band running at? Is it at the expected 72 kDa?

In order to use a blocking peptide correctly, the peptide needs to be incubated with the antibody in 5 - 10 times molar excess. Using both the antibody and the peptide at the same concentration will not allow proper blocking. The blocking assay looked like it worked though for the HEK lanes in the WB. Have you confirmed the empty vector since this band is blocked and is therefore considered specific?

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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