FITC Rat Anti-Mouse CD21/CD35
Clone 7G6 (RUO)
- Brand BD Pharmingen™
- Alternative Name CR2/CR1
- Concentration 0.5 mg/ml
- Isotype Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
Fluorescence microscopy (Reported)
- Immunogen Purified Mouse CR1
- Entrez Gene ID 12902
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 7G6 antibody recognizes an epitope shared by 145-150-kDa and 190-kDa complement receptor proteins, originally designated CR2 (CD21) and CR1 (CD35), respectively. In the mouse, CD21 and CD35 are expressed on the majority of peripheral B lymphocytes, on the majority of resident peritoneal macrophages and mast cells, on peripheral blood granulocytes after treatment with N-formyl-Met-Leu-Phe, and on follicular dendritic cells, but not on thymocytes, T cells, erythrocytes, or platelets. CD21 is a ligand-binding component of the CD19/CD21/CD81 signal-transduction complex associated with the antigen receptor on B lymphocytes. CD21/CD35 also co-localizes with CD19 on the surface of peritoneal mast cells. Cr2null mice display impaired inflammatory and humoral immune responses in vivo. The 7G6 mAb has been reported to inhibit rosette formation by C3d-bearing sheep erythrocytes, to block the complement dependent trapping of immune complexes by follicular dendritic cells, and to down-regulate mouse CD21/CD35 expression upon in vivo application, thus inhibiting primary antibody responses to immunization. Co-stimulation of B-cell differentiation via Sepharose-coupled 7G6 antibody has also been observed. The 7G6 mAb recognizes an epitope on CD35 distinct from the epitope recognized by anti-mouse CD35, clone 8C12, and it does not block binding of 8C12 mAb to mouse CD35.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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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 FITC under optimum conditions, and unreacted FITC was removed.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
We have observed that the staining intensity of FITC-conjugated 7G6 mAb is severely reduced after fixation of stained leukocytes (≤ 1 hour with 1% formaldehyde). Therefore, one should not fix the stained cells prior to flow cytometry. We have found that freshly-isolated leukocytes and cell lines may wait for analysis in wash buffer at 4°C, without fixation, for up to 18 hours post-staining without loss of viability. Activated lymphocytes may lose viability rapidly, and data should be collected within 5 hours post-staining.