Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

Immunofluorescence staining of CD74 in sections of formalin-fixed paraffin-embedded mouse spleen (positive) and mouse kidney (negative).

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323245 at 1/100 dilution and then incubated for 1 hour with ab200622 Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

ab323246 staining CD34 in NIH/3T3 (positive) and TRA-differentiated Neuro2a (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4° C with ab323246 at 5ug/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab200622 Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

Immunofluorescence staining of CD34 in sections of formalin-fixed paraffin-embedded mouse lung (positive) and mouse liver (negative).

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323246 at 1/100 dilution and then incubated for 1 hour with ab200622 Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

ab323245 staining CD74 in LPS treated Raw264.7 (positive) and TRA-differentiated Neuro2a (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4° C with ab323245 at 5ug/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab200622 Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

IHC image of human IgG1 H&L staining in a section of frozen normal human tonsil*.

The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab200622 at 1/5000 dilution (shown in green) and counterstained using ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Alexa Fluor® 488) (AB200622)

Negative IHC image of human IgG1 H&L staining in a section of frozen normal human cerebral cortex*.

The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab200622 at 1/5000 dilution (shown in green) and counterstained using ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.