Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199093 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

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

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MAPK14 KO HEK293T (human MAPK14 knock out human embryonic kidney epithelial cell, Left)/Parental HEK293T (Right) cells labelling p38 alpha/MAPK14 with ab316872 at 1/50 dilution (1 ug)/(Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) treated with 1mM Pervanadate for 30min (Red) / Untreated Jurkat (Green) cells labelling CD3 zeta (phospho Y142) with ab310016 at 1/500 dilution (0.1 ug) /(Red and Green) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labelling Integrin alpha 4/CD49D with ab316939 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

human PBMC are co-stained with CD3 conjugated Alexa Fluor®488.
Gated on viable cell.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling TREM1 with ab316943 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD3 conjugated Alexa Fluor®488.
Gated on viable cell.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD11a with ab317290 at 1/5000 dilution (0.01ug)/ Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Gated on viable cells. Cells are co-stained with CD3 conjugated to Alexa Fluor® 488.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 1%FBS for 3hours then 100nM Calyculin A for 10min (Red) / Untreated control (Dotted Red) cells labelling IKB alpha (phospho S36) with ab317485 at 1/5000 dilution (0.01ug) /Red and Dotted Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD45 with ab315960 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Negative control : 293T. Gated on viable cells.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD18 with ab316944 at 1/500 dilution (0.1ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD4 conjugated BV421.
Gated on viable cell.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / 293T cells transfected with a mouse CCR4 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling CCR4 with ab324334 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Cells were surface stained with isotype control or our antibody. Then fixed with 2% PFA for 10min followed by intracellularly stained with anti-myc tag conjugated to Alexa Fluor® 488.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a mouse CD27 expression vector (Middle) / 293T cells transfected with an empty expression vector (Right) cells labelling CD27 with ab324178 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were co-stained with Myc tag conjugated to Alexa Fluor®488.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling T-bet / Tbx21 with ab315957 at 1/5000 dilution (0.01ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Human PBMC are co-stained with CD19 conjugated to PE/Cy7, showing a distinct CD19 positive population with no cross reactivity to TBX21 positive cells.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 µg/ml PHA for 18h (bottom), either stained with ab326136 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 µg/ml human IgG, 10% normal goat serum and BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (1/10) to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab326136) (1x 106 in 100µl at 0.2 µg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). The cells were simultaneously stained with CD3. Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing left wild-type HAP1 positive cells and right negative FASN knockout HAP1 stained with ab319690 (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 (ab319690) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500 dilution)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor^® 647 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 40 mW Red laser (638nm) and 660/10 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) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HEK293T (green line) and HK1 knockout HEK293T stained with ab197864 (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 (ab197864) (1x 10⁶ in 100μl at 5 μg/ml (1/100)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HEK293T WT cells (black line) and HEK293T-HK1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa(human cervical adenocarcinoma epithelial cell) cells labelling JNK1 + JNK2 + JNK3 with ab315842 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag (upper) / 293T cells transfected with an empty vector containing a His tag (low) cells labelling RFP with ab323758 at 1/500 dilution (0.1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Cells are co-stained with Myc tag conjugated to Alexa Fluor®488.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of K-562 (human chronic myelogenous leukemia lymphoblast, Left)/Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling Integrin alpha 4/CD49D with ab316939 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cell.
Negative control : K-562 (PMID : 16105875).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab198587 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab198587, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor^® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte, Left) and Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD3D with ab198937 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor^® 647) (ab199093) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Negative control : THP-1.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) treated with 500ng/ml ionomycin and 10ng/ml PMA for 3 hours (Lower left and right) / Untreated control (Upper left and right) cells labelling CD69 with ab325108 at 1/500 dilution (0.1ug) / Upper right and Lower right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were stained with anti-CD4 conjugated to Brilliant Violet 421.
Gated on viable lymphoid Cells.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized hela(human cervical adenocarcinoma epithelial cell) cells labelling Ataxin 3 with ab315423 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Isotype (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with Myc-tagged ENPP3 overexpression construct (Middle) / 293T transfected with an empty vector containing a myc-His tag (Right) cells labelling ENPP3/B10 with ab319126 at 1/500 dilution (0.1ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody) (Blue).

Cells were co-stained with anti-myc conjugated to Alexa Fluor 488.
Gated on viable cells.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing left wild-type HAP1 positive cells and right negative ACLY knockout HAP1 stained with ab205430 (red 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 (ab205430) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 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)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype / 293T (human embryonic kidney epithelial cell) transfected with a TRDC (human TCR delta) protein expression vector containing a myc-His tag(Up Right) / 293T transfected with a TRGC1 protein (human TCR gamma) expression vector(Down Left) / 293T transfected with an empty vector containing a myc-His tag(Down Right) cells labelling TCR delta with ab317716 at 1/500 dilution (0.1ug)/ Up Right, Down Left and Down Right (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Hela (human cervical adenocarcinoma epithelial cell, Left)/U-937 (human histiocytic lymphoma monocyte, Right) cells labelling CD18 with ab316944 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cell.
Negative control : HeLa

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) / HEL (human erythroleukemia erythroblast, Right) cells labelling VEGF Receptor 3 with ab313899 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : MCF7. Gated on viable cells.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast) cells labelling IL-10 with ab305044 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / Daudi (human Burkitt's lymphoma lymphoblast, Middle) / K-562 (human chronic myelogenous leukemia lymphoblast, Right) cells labelling MICB with ab325223 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cells.
Negative control : 293T, Daudi (PMID : 28154561).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and PRDX2 knockout HAP1 stained with ab197041 (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 (ab197041) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500 dilution)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor^® 647 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 40 mW Red laser (638nm) and 660/10 bandpass filter.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Huh7 (human hepatocellular carcinoma epithelial cell) treated with 300nM thapsigargin for18hours (Red) / Untreated Huh7 (Dotted Red) cells labelling XBP1 with ab317358 at 1/5000 dilution(0.01ug)/Red and Dotted Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CLEC12A with ab317063 at 1/5000 dilution (0.01ug)/ Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Gated on viable cells. Cells were co-stained with anti-CD3 conjugated to Alexa Fluor® 488.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-87 MG(human glioblastoma-astrocytoma epithelial cell) cells labelling Ataxin 3 with ab315423 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HeLa (green line) and GYS1 knockout HeLa stained with ab326127 (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 (ab326127) (1x 106 in 100µl at 0.008 µg/ml (1/62500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HeLa WT cells (black line) and HeLa-GYS1 KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.This antibody gave a positive signal in wild-type HeLa fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labelling ILT-4 with ab313898 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Cells are co-stained with CD14 conjugated to BV510. Gated on viable cells.

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labelling KMT6 / EZH2 with ab317360 at 1/500 dilution(0.1ug)/(Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).