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BV421 Rat Anti-Mouse Axl
BV421 Rat Anti-Mouse Axl

Flow cytometric analysis of Axl expression on mouse cell lines.

        Left Plot: NIH3T3 (ATCC CRL-1658) mouse embryonic fibroblasts were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse Axl (Cat. No. 748028; solid line histogram) at 1.0 µg/test.

        Right Plot: ST-2 (DSMZ ACC 333) mouse bone marrow stromal cells were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse Axl (Cat. No. 748028; solid line histogram) at 1.0 µg/test.

        Cell Dissociation Buffer (Gibco™ 13151014) was used to detach the cell lines before staining. The fluorescence histograms showing Axl [or Ig Isotype control] staining were derived from gated events with the forward and side light-scatter characteristics of viable singlet cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

 

The above is qualification data only and does not represent a specific OptiBuild™ lot.

Flow cytometric analysis of Axl expression on mouse cell lines.

        Left Plot: NIH3T3 (ATCC CRL-1658) mouse embryonic fibroblasts were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse Axl (Cat. No. 748028; solid line histogram) at 1.0 µg/test.

        Right Plot: ST-2 (DSMZ ACC 333) mouse bone marrow stromal cells were stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD OptiBuild™ BV421 Rat Anti-Mouse Axl (Cat. No. 748028; solid line histogram) at 1.0 µg/test.

        Cell Dissociation Buffer (Gibco™ 13151014) was used to detach the cell lines before staining. The fluorescence histograms showing Axl [or Ig Isotype control] staining were derived from gated events with the forward and side light-scatter characteristics of viable singlet cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

 

The above is qualification data only and does not represent a specific OptiBuild™ lot.

Product Details
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BD OptiBuild™
Ark; adhesion-related kinase; Tyro7; Ufo; tyrosine-protein kinase receptor UFO
Mouse (Tested in Development)
Rat IgG2a, κ
Mouse Axl (Ala19–Trp445) Recombinant Protein
Flow cytometry (Qualified)
0.2 mg/ml
AB_2872489
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
748028 Rev. 2
Antibody Details
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175128

The 175128 monoclonal antibody specifically recognizes Axl which is also known as Adhesion related kinase (Ark), Tyro7, or Ufo. Tyro-3, Axl, and Mer constitute the TAM subfamily of receptor tyrosine kinases (RTK). Axl is a single-pass type I transmembrane glycoprotein comprised of an extracellular region with two immunoglobulin (Ig)-like domains and two fibronectin type III (FNIII) domains, a transmembrane segment and a conserved intracellular tyrosine kinase domain. Axl is variably expressed by multiple cell types including monocytes, macrophages, dendritic cells, platelets, endothelial cells, vascular smooth muscle cells, and fibroblasts. Axl binds to the vitamin K-dependent Growth arrest-specific protein 6 (Gas6) through its extracellular Ig-like domains. Ligand binding leads to receptor dimerization, and autophosphorylation of tyrosine residues within the cytoplasmic Axl domains. This results in the activation of downstream signaling pathways that control cellular adhesion, aggregation, phagocytosis/efferocytosis, proliferation, survival, and migration. Through these activities, Axl plays major roles including development, the regulation of hematopoiesis and immunity, and ensuring the integrity of the vascular system. Abnormal expression of Axl has been observed in various cancers and by tumor cell lines.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

748028 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
748028 Rev.2
Citations & References
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Development References (3)

  1. Aguilera TA, Rafat M, Castellini L, et al. Reprogramming the immunological microenvironment through radiation and targeting Axl.. Nat Commun. 2016; 7:13898. (Clone-specific: Flow cytometry). View Reference
  2. Paolino M, Choidas A, Wallner S, et al. The E3 ligase Cbl-b and TAM receptors regulate cancer metastasis via natural killer cells.. Nature. 2014; 507(7493):508-12. (Clone-specific: Flow cytometry). View Reference
  3. Rescigno J, Mansukhani A, Basilico C. A putative receptor tyrosine kinase with unique structural topology.. Oncogene. 1991; 6(10):1909-13. (Biology). View Reference
748028 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.