Rabbit IgG, monoclonal [EPR25A] - Isotype Control

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab133557 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab133557 at 1/60 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

Conjugated versions are available for this clone : Alexa Fluor^® 488 (ab199091), Alexa Fluor^® 647 (ab199093), R-PE (ab209478), APC (ab232814).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric overlay histogram analysis of 4% paraformaldehyde cells labelling CD276 KO HEK293T (ED040783) (Magenta and grey) cells permeabilised with 90% methanol and Parental HEK293T (EDWT04) (Green and black) cells permeabilised with 90% methanol labelling CD276 with ab227679 at 1/20 dilution (Magenta and Green) compared with a Rabbit monoclonal IgG (ab172730) (Black and Grey) - Isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150081), at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and ACACA knockout HAP1 stained with ab269272 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab269272) (1x 10⁶ in 100μl at 0.04 μg/ml (1/13975 dilution)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-ACACA KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) (Green and black) and KEAP1 KO HAP1(HO010289) (Magenta and grey) cells labelling with ab326338 at 1/500 dilution (0.1ug), compared with a Rabbit monoclonal IgG (ab172730) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human neutrophils cells labelling Cathelicidin/CLP with ab325356 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry (intracellular) analysis of transfected 293T cells. Cells were fixed with 4% PFA for 10 minutes and permeabilised with 90% methanol before staining for Prosurfactant Protein C in 293T cells transfected with a mouse SFTPC expression vector containing a myc-His-tag® using antibody ab270521 at 1/5000 dilution (0.01 ug) (middle panel). Staining was compared with 293T cells transfected with an empty expression vector containing a myc-His-tag® (right panel). and with an isotype control, rabbit monoclonal IgG (ab172730) (left panel). Goat anti‑rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody at a 1/5000 dilution. Cells were co-stained with anti-myc conjugated to Alexa Fluor®647.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and ACACA knockout HAP1 stained with ab269273 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab269273) (1x 10⁶ in 100μl at 0.008 μg/ml (1/74750 dilution)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-ACACA KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling LATS1/WARTS with ab243656 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC isolated CD14+ monocytes treated with 1000IU/ml IFNα for 24hours (Lower left and right) labelling Sialoadhesin/CD169 with ab183356 at 1/50 dilution (1ug)/ (Right) compared with a Rabbit monoclonal IgG (ab172730) / Left - Isotype Control (Left). Untreated Human PBMC isolated CD14+ monocytes (Upper left and right).

Cells blocked with 20ug/ml Human IgG.

Cells were surface stained with anti-CD14 conjugated to BV421. Then fixed with 4% PFA for 10 min followed by intracellular staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 10uM MG-132 for 4 hours (Green) and Untreated control (Magenta) cells labelling Nrf2 with ab325240 at 1/500 dilution (0.1ug) / Green and Red compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell, Left) THP-1 (human monocytic leukemia monocyte, Right) cells labelling with ab325712 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor®488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : 293T.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram of 4% Paraformaldehyde MG 63 (human osteosarcoma fibroblast) cells permeabilised with 90% methanol labelling RBPJK with ab180588 at 1/50 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150081) , at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) (Green and black) and ANXA5 KO HAP1(HO010326) (Magenta and grey) cells labelling Annexin V/ANXA5 with ab325388 at 1/500 dilution (0.1ug) / Green and magenta compared with a Rabbit monoclonal IgG (ab172730) / Black and Grey isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry (intracellular) analysis of transfected 293T cells. Cells were fixed with 4% PFA for 10 minutes and permeabilised with 0.1% Tween-20 before staining for Langerin in 293T cells transfected with a human Langerin expression vector containing a myc-His-tag® using ab283686 1/1000 dilution (0.1ug) conjugated to Alexa Fluor® 647. (Middle panel). Staining was compared with 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right panel) and with an isotype control, Rabbit monoclonal IgG (ab172730) (left panel). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody at a 1/5000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram of 4% paraformaldehyde HEK-293 (human embryonic kidney epithelial cell) cells permeabilised with 90% methanol labelling Vinculin with ab196454 at 1/500 dilution (Red line) compared with a Rabbit monoclonal IgG (ab172730) as Isotype control (Black line) and an unlabelled control (cells without incubation with primary antibody) (Blue line).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) (Green and black) IL17RA KO HAP1(HO010217) (Magenta and grey) cells labelling IL-17RA Receptor with ab325606 at 1/500 dilution (0.1ug) / Magenta and Green compared with a Rabbit monoclonal IgG (ab172730) / Black and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram showing wild-type A-549 (green line) and SHMT2 knockout A-549 stained with ab316328 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab316328) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in A-549 WT cells (black line) and A-549-SHMT2 KO cells (grey line), at the same conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized 293T cells transfected with a mouse Cd244 expression vector containing a myc-His-tag® (Middle) 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling 2B4 with ab325752 at 1/50000 dilution (0.001ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were surface stained with isotype control or our antibody. Then fixed with 4% PFA for 10min followed by intracellularly stained with anti-myc tag conjugated to Alexa Fluor® 647.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram of WERI-Rb-1 (human Eye Retina grape-like clusters of round cells) labelling GPC2 with ab324378 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Secondary antibody details, Goat anti-Rabbit IgG (Alexa Fluor^® 647 (ab150083), at 1/5000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human purified neutrophils cells labelling IL-17RA Receptor with ab325606 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CXCR2 with ab325787 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with CD56 conjugated to Brilliant Violet 421. Gated on viable lymphocyte cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD5 with ab325643 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Cells were co-stained with anti human CD3 conjugated to Alexa Fluor®647.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human neutrophils cells labelling CD66b with ab325777 at 1/5000 (0.01ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Unstimulated HL-60 (Human acute promyelocytic leukemia promyeloblast) (Lower) HL-60 stimulated with 1.25% DMSO for 5 days (Upper) cells labelling CD66b with ab325777 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of LS513 (human Large intestine Cecum epithelial cell, Right) Daudi (human Burkitt's lymphoma lymphoblast, Left) cells labelling Tspan-8 with ab326570 at 1/50 dilution (1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : Daudi.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CXCR2 with ab325787 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with CD19 conjugated to PE/CY7. Gated on viable lymphocyte cells.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell, Left ) / PC-3 (human prostate adenocarcinoma epithelial cell, Right ) cells labelling FOXA1 + FOXA2 + FOXA3 with ab317046 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : HCT 116

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling KMT6 / EZH2 (phos Thr487) with ab300567 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) treated with 20µM nocodazole for 24 hours (Right) / Untreated control (Left) cells labelling KMT6 / EZH2 (phos Thr487) with ab300567 at 1/500 dilution (0.1ug)/ Left and Right (Red) compared with an isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of MOLT-4 (human lymphoblastic leukemia T lymphoblast) (Right) HeLa (human cervical adenocarcinoma epithelial cell) (Left) cells labelling CD5 with ab325643 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Negative control : HeLa (PMID : 15187131).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a human PLCZ1 expression vector containing a myc-His-tag® (Middle) 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling PLCZ1 with ab325528 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling with ab326720 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable lymphoid cells. Cells were co-stained with CD3 conjugated to Brilliant Violet 421.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human purified neutrophils cells labelling CXCR2 with ab325787 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC isolated CD14+ monocytes treated with IFNa (1000 IU/ml ) for 24 hours (Lower left and right) Human PBMC (human peripheral blood mononuclear cell) isolated CD14+ monocytes (Upper left and right) cells labelling Sialoadhesin/CD169 with ab326438 at 1/50 dilution (1ug) / Upper right and Lower right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Cells were co-stained with anti-CD14 conjugated to Brilliant Violet 421.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SU-DHL-1 (human histiocytic lymphoblast-like cell, Left) and karpas-299 (human T cell lymphoma cell, Right) cells labelling ROR gamma with ab325651 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : SU-DHL-1(PMID : 29588546).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) SH-SY5Y (human neuroblastoma epithelial cell, Left) cells labelling with ab326720 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Negative control : SH-SY5Y.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Daudi (human Burkitt's lymphoma lymphoblast, Left) TF-1 (human erythroleukemia erythroblast, Right) cells labelling Leptin Receptor with ab325970 at 1/50 dilution (1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : Daudi.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling with ab326720 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable lymphoid cells. Cells were co-stained with CD4 conjugated to APC/Fire™ 750.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram of Negative control : HeLa (human cervical adenocarcinoma epithelial cells, Left) and IMR-32 (human neuroblastoma neuroblasts, Right) labelling GPC2 with ab324378 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Secondary antibody details, Goat anti-Rabbit IgG (Alexa Fluor^® 647 (ab150083), at 1/5000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometric analysis of MOLT-4 (human lymphoblastic leukemia T lymphoblast, Left) THP-1 (human monocytic leukemia monocyte, Right) cells labelling IL-17RA Receptor with ab325606 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : MOLT-4.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB172730)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and VDAC1 knockout HAP1 stained with ab154856 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab154856) (1x 10⁶ in 100μl at 0.2 μg/ml (1/195 dilution)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-VDAC1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.