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BB515 Rat Anti-Mouse CD370 (Clec9A)
BB515 Rat Anti-Mouse CD370 (Clec9A)
Multicolor flow cytometric analysis of CD370 (Clec9A) expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091), BD Horizon™ BUV737 Rat Anti-Mouse CD8a (Cat. No. 564297), BD Horizon PE-CF594 Hamster Anti-Mouse CD11c (Cat. No. 562454) antibodies and either BD Horizon BB515 Rat IgG2a κ Isotype Control (Cat. No. 564418; Left Panel) or BD Horizon BB515 Rat Anti-Mouse CD370 (Clec9A) antibody (Cat. No. 565320/566033; Right Panel). Two color flow cytometric dot plots showing the correlated expression of CD11c and CD370 (Clec9A) [or Ig Isotype control staining] were derived from CD4- CD8a+ gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of CD370 (Clec9A) expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091), BD Horizon™ BUV737 Rat Anti-Mouse CD8a (Cat. No. 564297), BD Horizon PE-CF594 Hamster Anti-Mouse CD11c (Cat. No. 562454) antibodies and either BD Horizon BB515 Rat IgG2a κ Isotype Control (Cat. No. 564418; Left Panel) or BD Horizon BB515 Rat Anti-Mouse CD370 (Clec9A) antibody (Cat. No. 565320/566033; Right Panel). Two color flow cytometric dot plots showing the correlated expression of CD11c and CD370 (Clec9A) [or Ig Isotype control staining] were derived from CD4- CD8a+ gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
製品詳細
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BD Horizon™
CD370; CLC9A; Clec9a; DNGR-1; C-type lectin domain family 9 member A
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Mouse Clec9A Peptide
Flow cytometry (Routinely Tested)
0.2 mg/ml
232414
AB_2739179
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™ BB515 under optimum conditions and unconjugated antibody was removed.

推奨アッセイ手順

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

関連製品

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.1 mg カタログ番号 553141
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Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.5 mg カタログ番号 553142
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Brilliant Stain Buffer RUO
サイズ 1000 Tests カタログ番号 566349
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Brilliant Stain Buffer Plus RUO
サイズ 1000 Tests カタログ番号 566385
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Stain Buffer (FBS) RUO
サイズ 500 mL カタログ番号 554656
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Stain Buffer (BSA) RUO
サイズ 500 mL カタログ番号 554657
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565320 Rev. 3
抗体の詳細
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10B4

The 10B4 monoclonal antibody specifically binds to mouse Clec9A. Mouse Clec9A (C-type lectin domain family member 9A) is also known as DNGR1 (Dendritic cell natural killer lectin group receptor 1). It is a type II membrane protein with a single extracellular C-type lectin domain. Clec9A is a dendritic cell subtype-restricted C-type lectin-like receptor. Clec9A is selectively expressed on plasmacytoid dendritic cells and CD8+ myeloid dendritic cells. Clec9A reportedly serves as a receptor for necrotic cells. It can mediate the cross-presentation of dead-cell associated antigens in a Syk-dependent manner.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.        

565320 Rev. 3
フォーマットの詳細
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
565320 Rev.3
引用&参考文献
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View product citations for antibody "565320" on CiteAb

Development References (3)

  1. Caminschi L, Proietto AL, Ahmet F, et al. The dendritic cell subtype restricted C-type lectin Clec9A is a target for vaccine enhancement. Blood. 2008; 112(8):3264-3273. (Immunogen: Flow cytometry, Functional assay, Immunohistochemistry, In vivo exacerbation). View Reference
  2. Huysamen C, Willment JA, Dennehy KM, Brown GD. CLEC9A is a novel activation C-type lectin-like receptor expressed on BDCA3+ dendritic cells and a subset of monocytes. J Biol Chem. 2008; 283(24):16693-16701. (Biology). View Reference
  3. Sancho D, Joffre OP, Keller AM, et al. Identification of a dendritic cell receptor that couples sensing of necrosis to immunity. Nature. 2009; 458(7240):899-903. (Biology). View Reference
565320 Rev. 3

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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.