Key Product Details

Species Reactivity

Validated:

Human

Cited:

Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry

Cited:

Immunocytochemistry

Label

Biotin

Antibody Source

Monoclonal Mouse IgG2B Clone # 5D3
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Product Specifications

Immunogen

3T3 cells transduced with human ABCG2

Specificity

Detects human ABCG2.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human ABCG2 Biotinylated Antibody

ABCG2 in A549 Human Cell Line.

ABCG2 was detected in immersion fixed A549 human lung carcinoma cell line using Mouse Anti-Human ABCG2 Biotinylated Monoclonal Antibody (Catalog # BAM995) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human ABCG2 Biotinylated Antibody

Application
Recommended Usage

Flow Cytometry

2.5 µg/106 cells
Sample: Human MCF-7 cell line

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed A549 human lung carcinoma cell line

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ABCG2

Hematopoietic stem cells are known to express a membrane transporter molecule, known as P-glycoprotein (Pgp), that is encoded by the multidrug resistance gene 1 (MDR1) (1, 2). Expression of Pgp appears to confer a proliferative advantage to stem cells through its anti-apoptotic effects (3, 4). An additional transporter molecule known as ABCG2 (ATP-binding cassette gene 2) or Bcrp1 (Breast cancer resistance protein 1), first identified in a breast cancer cell line (5), is expressed on stem cells (6). ABCG2 belongs to a family of molecules that span the cell membrane six times and can exist as either homo or hetero dimers linked by a short intracellular flexible linker region that plays an important role in the efflux of a wide range of substrates (7, 8). Although these transporter molecules have initially been thought to play a role in drug resistance, they have been found to have utility in better characterizing primitive stem cells. For example, the “side-population” of hematopoietic stem cells, characterized by their inability to retain high levels of the intracellular staining dyes Hoechst 33342 and Rhodamine 123, has been found to express high levels of ABCG2. Of interest is the observation that ABCG2 function has been linked to the efflux of the Hoechst dye (6). Furthermore, there is now evidence that this monoclonal can be used as a cell surface marker to identify hematopoietic stem cells within the bone marrow fraction of lineage negative cells (6). The expression of ABCG2 appears greatest on CD34- cells and is downregulated with the acquisition of CD34 on the cell surface (6).

References

  1. Chaudhary, P.M. and I.B. Roninson (1991) Cell 66:85.
  2. Sorrentino, B.P. et al. (1995) Blood 86:491.
  3. Pallis, M. and N. Russell (2000) Blood 95:2897.
  4. Johnstone, R.W. et al. (1999) Blood 93:1075.
  5. Doyle, L.A. et al. (1998) Proc. Natl. Acad. Sci. USA 95:15665.
  6. Zhou, S. et al. (2001) Nat. Medicine 7:1028.
  7. Hrycyna, C.A. et al. (1998) Biochem. 37:13660.
  8. Bunting, K.D. (2002) Stem Cells 20:11.

Long Name

ATP-binding Cassette Transporter G2

Alternate Names

ABCP, BCRP, CD338, MXR

Entrez Gene IDs

9429 (Human)

Gene Symbol

ABCG2

Additional ABCG2 Products

Product Documents for Human ABCG2 Biotinylated Antibody

Certificate of Analysis

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Product Specific Notices for Human ABCG2 Biotinylated Antibody

For research use only

Citations for Human ABCG2 Biotinylated Antibody

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Protocols

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