Use negative controls (unstained and
isotype control antibodies) to determine gating of populations.
HL - 60 cells were stained with 1 µg / mL ab14715 (blue) or an equal amount of
an isotype control antibody (red) and analyzed by flow cytometry.
An isotype control for the secondary antibody was used as a negative control in both samples.
(d) Histogram of membrane - bound Tie2 expression in HUVEC and BP compared to
isotype control measured by flow cytometry.
After washing with PBS and blocking for 30 minutes (IHC / ICC Blocking Buffer - Low Protein; eBioscience), cells were incubated with anti - Oct ‑ 4 antibody (diluted 1:150; eBiosicence) or an IgG2a K
isotype control (diluted 1:150; eBiosicence) for one hour.
Isotype control antibody (black line) was rabbit IgG (monoclonal)(1μg / 1x106 cells) used under the same conditions.
Unstained,
isotype control, and E-cadherin (E-cad)- stained cells are shown in the histogram.
Rat anti-mouse ST2 - blocking Ab and mouse IgG1
isotype control Ab were provided by Amgen.
An isotype control IgG was run in parallel and showed no positive staining (not shown here).
Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg / 1x106 cells) used under the same conditions.
(C) Survival of gp33 - immunized mice with LCMV infection following 30 d rest period compared with control mice with ST2 blockade or
isotype control Ab treatment.
LCMV - infected mice were injected i.p. with 150 μg ST2 blocking Ab or
isotype control every other day beginning on day 2 postinfection (14).
Rat anti-mouse IFN - γ (XMG1.2, 0.5 mg), rat anti-mouse TNF - α (XT3.11, 1 mg), and rat IgG2a
isotype control (2A3, 0.5 mg) were injected i.p. every third day beginning on day 2 postinfection (8).
(E) Mice were treated beginning day 2 postinfection, and every 3 d thereafter with
isotype control, IFN - γ blockade only, TNF - α blockade only, or IFN - γ / TNF - α dual blockade following infection.
(a) Top panel: Unstained,
isotype control, and ESA - stained cells are shown in the histogram.
Prf1 − / − mice were immunized against gp33 or with control procedure, rested for 30 d, infected with LCMV, and treated with either IFN - γ blockade or
isotype control beginning day 2 postinfection, and every 3 d thereafter.
(B) gp33 - immunized and control mice were treated with IFN - γ blockade or
isotype control Ab treatment.
They were then washed in FACS buffer I and probed in FACS buffer I containing a 1:500 dilution of r - phycoerythrin - conjugated goat anti-mouse IgG (Caltag Laboratories, Carlsbad, CA) or an appropriate
isotype control for 15 min in the dark at 4 °C.
Cells were stained with STAT6 monoclonal antibody or mouse IgG1
isotype control antibody or without the primary antibody.
Inhibition of cell proliferation (B) of MCF7, MCF7 / HER2, or BT474 cells treated for 6 days with F3 - IgG, trastuzumab, or
isotype control antibody.
The isotype control at 1 ug / mL shows no higher signal than the no primary negative control.
A non-specific
isotype control antibody (human IgG1) showed no reactivity.
An isotyping ELISA was performed by coating a 96 - well plate with 1.25 ug / mL of the IgG1
Isotype Control Antibody and detecting with Alexa Fluor conjugates specific to mouse IgG1, IgG2a, IgG2b, IgG3, IgM and heavy and light chains (H&L) of IgG.
Not exact matches
Quadrants were set based on
isotype - stained
controls.
Breast carcinoma cells (5 × 103 cells / well) were seeded into a 96 - well plate and treated with trastuzumab,
isotype - matched
control IgG, or F3 - IgG at concentrations ranging from 0.01 to 10 µg / mL.
To assess the ability of F3 - IgG to inhibit the proliferation of HER2 - overexpressing breast carcinoma cells, we measured the proliferation of BT474, MCF7 / HER2 and MCF7 breast cancer cell lines in the presence of trastuzumab, F3 - IgG, or an
isotype - matched
control antibody.