Anti-STAT2 antibody [Y141] (ab32367)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y141] to STAT2
- Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-STAT2 antibody [Y141]
See all STAT2 primary antibodies -
Description
Rabbit monoclonal [Y141] to STAT2 -
Host species
Rabbit -
Specificity
This antibody detects both long and short forms of STAT2. -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanWB MouseRatHuman -
Immunogen
Synthetic peptide within Human STAT2 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: A549, K562, HAP1, Ramos, HeLa and THP1 cell lysates; Mouse and Rat brain tissue lysates ICC/IF: THP-1 cells IHC-P: Human hepatocellular carcinoma and thyroid cancer tissue, Mouse and Rat liver Tissue Flow Cyt: Ramos cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y141 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : STAT2 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab32367 was shown to react with STAT2 in A549 wild-type cells in western blot with loss of signal observed in STAT2 knockout cell line ab267004 (STAT2 knockout cell lysate ab257183). Wild-type and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk before incubation with ab32367 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : STAT2 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32367 was shown to react with STAT2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267005 (knockout cell lysate ab257184) was used. Wild-type A549 and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STAT2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32367 was shown to react with STAT2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261819 (knockout cell lysate ab257182) was used. Wild-type HeLa and STAT2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : STAT2 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32367 Anti-STAT2 antibody [Y141] was shown to specifically react with STAT2 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267006 (knockout cell lysate ab257185) was used. Wild-type and STAT2 knockout samples were subjected to SDS-PAGE. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-STAT2 antibody [Y141] (ab32367)
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : STAT2 knockout HAP1 whole cell lysate
Lane 3 : K562 whole cell lysate
Lane 4 : THP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 98 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32367 (unpurified) observed at 97 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab32367 was shown to recognize STAT2 in wild-type HAP1 cells as signal was lost at the expected MW in STAT2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and STAT2 knockout samples were subjected to SDS-PAGE. Ab32367 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling STAT2 with purified ab32367 at 1/60 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of THP-1 (human monocytic leukemia monocyte) cells labeling STAT2 with purified ab32367 at 1/50 dilution (10 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
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All lanes : Anti-STAT2 antibody [Y141] (ab32367) at 1/1000 dilution (Purified)
Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : Mouse brain lysates
Lane 4 : Rat brain lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 113,97 kDa why is the actual band size different from the predicted?Blocking/Diluting Buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemical analysis of paraffin-embedded human thyroid cancer using ab32367 (unpurified). Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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