Rabbit IgG, monoclonal [EPR25A] - Isotype Control

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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).

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A-673 (human muscle Ewings sarcoma cell line) treated with 300ng/ml BFA for 24 hours (Left) A375 (human malignant melanoma epithelial cell) treated with 300ng/ml BFA for 24 hours (Right) cells labelling IL-24 with ab325236 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.

Low expression : A-673 treated with 300ng/ml BFA for 24 hours

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

Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left)/ LNCaP (Human prostate carcinoma epithelial cell, Right) cells labelling TMPRSS2 with ab280567 at 1/500 dilution (0.1μg)/ red compared with 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^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody.

Negative control : HEK-239T (PMID : 20631123).

Gated on viable cells.

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

Negative control : Raji. (PMID : 11207281).

Raji (human Burkitt's lymphoma cell line) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)(Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ^® 488) at 1/2000 dilution.

Gated on viable cells.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RD (human muscle multinucleated spindle-shaped cell) (Right) U-937 (human histiocytic lymphoma monocyte) (Left) cells labelling Nestin with ab324884 at 1/50000 dilution (0.001ug) / 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 : U-937.

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

Flow cytometric analysis of Raji (Human Burkitt's lymphoma B lymphocyte, Left) / BxPC-3 (Human pancreas adenocarcinoma epithelial cell, Right) cells labelling ULBP2+ULBP5+ULBP6 with ab290731 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) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : Raji.Gated on viable cells.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells (human embryonic kidney epithelial cell) transfected with a fbpA expression vector containing a myc tag cells labelling Mycobacterium tuberculosis Ag85B with ab312328 at 1/500 dilution (0.1 ug)/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.

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

Flow cytometric analysis of MeWo (human malignant melanoma fibroblast, Right) HepG2 (human hepatocellular carcinoma epithelial cell, Left) cells labelling IGSF11 with ab325341 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.
Negative control : HepG2.

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

Flow cytometric analysis of 786-O (human kidney epithelial cell, Left) DLD-1 (human colorectal adenocarcinoma epithelial cell, Right) cells labelling LOX 1 with ab325339 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)/ Blue.

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

Gated on viable cells. Negative control : 786-O.

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

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD18 with ab325521 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 anti human CD14 conjugated to Alexa Fluor®647. Gated on viable cells.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NES KO HAP1(HO010139) (NES knockout human chronic myelogenous leukemia near-haploid cell) (Magenta) Parental HAP1(HOWT01) (Green) cells labelling Nestin with ab324884 at 1/50000 dilution (0.001ug) / Green and Magenta 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.

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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) cells labelling Fibrillarin with ab314506 at 1/500 dilution (0.1 ug)/Red (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

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

Flow cytometric analysis of Isotype (Left) / 293T cells transfected with a human SIGLEC6 expression vector containing a GFP-tag® (Middle) / 293T cells transfected with an empty expression vector containing a GFP-tag® (Right) cells labelling SIGLEC6 with ab323795 at 1/5000 dilution (0.01 ug).

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

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

Flow cytometric analysis of Isotype (Left) / 293T cells transfected with a SIGLEC6 expression vector containing a GFP-tag (Middle) /293T cells transfected an empty vector containing a GFP-tag (Right) cells labelling SIGLEC6 with ab317309 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Gated on viable cells.

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

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

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell) transfected with a human EGFRvIII expression vector containing a His-Myc tag (top left, bottom left); human wild-type EGFR expression vector containing a His-Myc tag (top right); empty vector containing a His-Myc tag (bottom right). Cells were fixed with 2% paraformaldehyde and permeabilised with 0.1% Tween-20. Cell labelling of EGFRvIII with ab313646 at 1/500 dilution. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730). No cross-reactivity with human wild-type EGFR.

Cells were surface stained with primary antibody. Fixed with 2% PFA for 10 min followed by intracellularly staining with anti-Myc tag conjugated to Alexa Fluor^® 647.

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

Flow cytometric analysis of SK-MEL-28 (human malignant melanoma cell, Right) THP-1 (human monocytic leukemia monocyte, Left) cells labelling IGSF11 with ab325341 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.
Negative control : THP-1.

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

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling BATF3 with ab302568 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 1/2000 dilution was used as the secondary antibody. Cells were stained with anti-CD11c conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab302568.

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

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling IL1 Receptor I/IL-1R-1 with ab325438 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

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

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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) cells labelling Histone H2A (symmetric di methyl R3) + Histone H4 (symmetric di methyl R3) with ab309354 at 1/500 dilution (0.1 ug)/Red (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

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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.

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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) 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 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.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human IL-24 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling IL-24 with ab325236 at 1/500 dilution (0.1ug) / 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 co-stained with Myc tag conjugated to Alexa Fluor®647.

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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.

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

Flow cytometric analysis of U937 (human histiocytic lymphoma monocyte) (Right) / HeLa (human cervical adenocarcinoma epithelial cell) (Left) cells labelling CD18 with ab325521 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.

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

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / 293T (human embryonic kidney epithelial cell) cells transfected with a human IL17A expression vector containing a Myc tag (Middle) / 293T cells transfected with an empty expression vector containing a Myc tag (Right) cells labelling IL17A with ab323803 at 1/5000 dilution (0.01 ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

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

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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).

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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.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human IL-7 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling IL-7 with ab324645 at 1/500 dilution (0.1ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Cells are co-stained with Myc tag conjugated to Alexa Fluor®647.

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

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A-172 (human brain glioblastoma) cells labelling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1/500 dilution (0.1ug) (Red) (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

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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.

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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.

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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.

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

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and PRDX2 knockout HAP1 stained with ab133481 (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 (ab133481) (1x 10⁶ in 100μl at 0.008 μg/ml (1/14375 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-PRDX2 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.

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

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and FASN knockout HAP1 stained with ab128870 (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 (ab128870) (1x 10⁶ in 100μl at 0.008 μg/ml (1/69250 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-FASN 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.

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

Flow cytometry overlay histogram showing left wild-type HAP1 positive cells and right negative ACLY knockout HAP1 stained with ab40793 (red 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 (ab40793) (1x 10⁶ in 100μl at 0.04 μg/ml (1/6225 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 (black line) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 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.

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

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and PRDX2 knockout HAP1 stained with ab109367 (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 (ab109367) (1x 10⁶ in 100μl at 0.04 μg/ml (1/21200 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-PRDX2 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.

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

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling IL1 Receptor I/IL-1R-1 with ab325438 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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 Alexa Fluor®647. Gated on viable cells.

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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.

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

Flow cytometric analysis of Jurkat (human T cell leukemia T lymphocyte, Left) / HUVEC (human umbilical vein endothelial cell, Right) cells labelling SREC-I with ab300407 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) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Negative control : Jurkat (PMID : 23892722)