BB515 Rat Anti-Mouse I-A/I-E

BD Horizon™ BB515 Rat Anti-Mouse I-A/I-E

Name: BB515 Rat Anti-Mouse I-A/I-E
Brand: BB515 Rat Anti-Mouse I-A/I-E
SKU: 565254

Clone 2G9 (RUO)

Two-color flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on Mouse splenocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. C57BL/6 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 CD45R/B220 (Cat. No. 553092/561880) and APC Rat Anti-Mouse CD11b (Cat. No. 553312/561690) antibodies, and either BB515 Rat IgG2a, κ Isotype Control (Cat. No. 564418; Left Panel), BD Horizon BB515 Rat Anti-Mouse I-A/I-E (Cat. No. 565254; Middle Panel), or FITC Rat Anti-Mouse I-A/I-E (Cat. No. 553623/562009; Right Panel). Two-color flow cytometric contour plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Mouse splenic leucocytes from SJL mice (H-2s haplotype) did not stain positively with the 2G9 antibody (data not shown). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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Catalog No:562060

List Price:Inquire

Unit:EA (1 Each)

Size:

0.1 mg

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Product Details

Brand: BD Horizon™

Alternative Name: H-2I; I-Ad, -Ed; I-Eb and I-Ek MHC class II alloantigens; Ia Ag; MHC II

Reactivity: Mouse (QC Testing)

Isotype: Rat DA, also known as DA/HA IgG2a, κ

Immunogen: BALB/c mouse epidermal Langerhans cells

Application: Flow cytometry (Routinely Tested), Immunofluorescence (Reported)

Concentration: 0.2 mg/ml

Entrez Gene ID: 14961, 14969

RRID: AB_2739134

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Alexa Fluor™ is a trademark of Life Technologies Corporation.
For U.S. patents that may apply, see bd.com/patents.

Data Sheets

Technical Data Sheet (TDS)

Companion Products

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.1 mg Cat No. 553141

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

Stain Buffer (FBS) RUO

Size 500 ML Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 ML Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Lysing Buffer RUO

Size 100 ML Cat No. 555899

View Details

565254 Rev. 4

Antibody Details

2G9

The 2G9 monoclonal antibody reacts with the I-Ad and I-Ed MHC class II alloantigens. It also reacts with transfectants expressing I-Eb and I-Ek and with cells from mice of the H-2p and H-2q haplotypes. This antibody is non-reactive with cells from SJL (H-2s) and NOD (H-2g7) mice. Flow cytometric analysis indicates that the 2G9 and M5/114.15.2 monoclonal antibodies have comparable reactivity on cells from mice with I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek alloantigens.

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.

565254 Rev. 4

Format Details

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.

Format: BB515

Excitation Max: 490 nm

Emission Max: 515 nm

565254 Rev.4

Citations & References

View Development References

Development References (4)

Becker D, Mohamadzadeh M, Reske K, Knop J. Increased level of intracellular MHC class II molecules in murine Langerhans cells following in vivo and in vitro administration of contact allergens. J Invest Dermatol. 1992; 99(5):545-549. (Immunogen: Immunofluorescence, Immunoprecipitation). View Reference
Farquhar CA, Paterson AM, Cobbold SP, et al. Tolerogenicity is not an absolute property of a dendritic cell. Eur J Immunol. 2010; 40(6):1728-1737. (Clone-specific: Flow cytometry). View Reference
Wculek SK, Amores-Iniesta J, Conde-Garrosa R, Khouili SC, Melero I, Sancho D. Effective cancer immunotherapy by natural mouse conventional type-1 dendritic cells bearing dead tumor antigen.. J Immunother Cancer. 2019; 7(1):100. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
Zeng Q, Liu JC, Yang C, et al. Gut microbiota-driven pathogenic Th17 cells mediated autoimmune epididymo-orchitis in a mouse model of colitis.. iScience. 2025; 28(5):112508. (Clone-specific: Immunofluorescence). View Reference

View All (4)

565254 Rev. 4

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.