Recombinant Anti-PTP1B antibody [EPR22474] (ab244207)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22474] to PTP1B
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PTP1B antibody [EPR22474]
See all PTP1B primary antibodies -
Description
Rabbit monoclonal [EPR22474] to PTP1B -
Host species
Rabbit -
Specificity
IHC is recommended for human only.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, HepG2, A549, MCF7, SW480, Jurkat, RAW 264.7, PC-12, NIH/3T3 and HCT 116 whole cell lysates. IHC-P: Human breast cancer and colon cancer tissue. ICC/IF: HeLa, wild type HAP1 and HCT 116 cells. Flow Cyt (intra): HeLa and HCT 116 cells. IP: PTP1B IP in HCT 116 whole cell lysate.
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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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22474 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab244207 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/50.
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WB |
1/1000. Predicted molecular weight: 50 kDa.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC is recommended for human only. |
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ICC/IF |
1/50.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/50. |
WB
1/1000. Predicted molecular weight: 50 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. IHC is recommended for human only. |
ICC/IF
1/50. |
IP
1/30. |
Target
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Function
May play an important role in CKII- and p60c-src-induced signal transduction cascades. -
Sequence similarities
Belongs to the protein-tyrosine phosphatase family. Non-receptor class 1 subfamily.
Contains 1 tyrosine-protein phosphatase domain. -
Post-translational
modificationsOxidized on Cys-215; the Cys-SOH formed in response to redox signaling reacts with the alpha-amido of the following residue to form a 4-amino-3-isothiazolidinone serine cross-link, triggering a conformational change that inhibits substrate binding and activity. The active site can be restored by reduction. -
Cellular localization
Endoplasmic reticulum membrane. - Information by UniProt
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Database links
- Entrez Gene: 5770 Human
- Entrez Gene: 19246 Mouse
- Entrez Gene: 24697 Rat
- Omim: 176885 Human
- SwissProt: P18031 Human
- SwissProt: P35821 Mouse
- SwissProt: P20417 Rat
- Unigene: 417549 Human
see all -
Alternative names
- PTP1B antibody
- Non receptor tyrosine phosphatase 1 antibody
- Protein phosphotyrosylphosphatase 1B antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human colon cancer (PMID:27752061) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PTPN1 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab244207 observed at 51 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab244207 was shown to react with PTP1B in wild-type HeLa cells in Western blot with loss of signal observed in PTPN1 knockout cell line ab265014 (PTPN1 knockout cell lysate ab257617). Wild-type HeLa and PTPN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab244207 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 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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All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PTPN1 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDaLanes 1- 2: Merged signal (red and green). Green - ab244207 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab244207 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265014 (CRISPR/Cas9 edited cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 CRISPR/Cas9 edited 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. ab244207 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 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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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cells is observed. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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PTP1B was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab244207 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab244207 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HCT 116 whole cell lysate 10 µg (Input).
Lane 2: ab244207 IP in HCT 116 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab244207 in HCT 116 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.The observed MW is consistent with what described in the literatures. (PMID: 18253097; PMID: 11895943; PMID: 19797268).
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All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PTP1B knockout HAP1 whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 50 kDaab244207 was shown to specifically react with PTP1B in wild-type HAP1 cells as signal was lost in PTP1B knockout cells. Wild-type and PTP1B knockout samples were subjected to SDS-PAGE. ab244207 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
The expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution
Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 50 kDaThe expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.
Blocking/Dilution buffer: NFDM/TBST.
Exposure time: 37 seconds.
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All lanes : Anti-PTP1B antibody [EPR22474] (ab244207) at 1/1000 dilution
Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDaThe expression profile observed is consistent with what has been described in the literature (PMID: 18253097; PMID: 11895943; PMID: 19797268). The bands below 50 kDa may represent truncated forms and cleaved fragments.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 48 seconds.
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Higher cytoplasmic expression in human breast cancer than that of adjacent normal tissues (PMID: 27465552) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HCT 116 cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Wild type and PTP1B-knockout HAP1 (Human chronic myelogenous leukemia near-haploid cell line) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing no staining in PTP1B-knockout HAP1 cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/50 (red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (7)
ab244207 has been referenced in 7 publications.
- Wiede F et al. PTP1B Is an Intracellular Checkpoint that Limits T-cell and CAR T-cell Antitumor Immunity. Cancer Discov 12:752-773 (2022). PubMed: 34794959
- Mo X et al. Tyrosine phosphorylation tunes chemical and thermal sensitivity of TRPV2 ion channel. Elife 11:N/A (2022). PubMed: 35686730
- Li Y et al. Hippocampal miRNA-144 Modulates Depressive-Like Behaviors in Rats by Targeting PTP1B. Neuropsychiatr Dis Treat 17:389-399 (2021). PubMed: 33603377
- Su C et al. Roux-en-Y Gastric Bypass Improves Hepatic Glucose Metabolism Involving Upregulation of Sirt1 in Type 2 Diabetes Mellitus. Diabetes Metab Syndr Obes 14:2269-2280 (2021). PubMed: 34045876
- Pan Z et al. UBE2D3 Activates SHP-2 Ubiquitination to Promote Glycolysis and Proliferation of Glioma via Regulating STAT3 Signaling Pathway. Front Oncol 11:674286 (2021). PubMed: 34195079
- Chen Q et al. TRIM18-Regulated STAT3 Signaling Pathway via PTP1B Promotes Renal Epithelial-Mesenchymal Transition, Inflammation, and Fibrosis in Diabetic Kidney Disease. Front Physiol 12:709506 (2021). PubMed: 34434118
- Liu X et al. A self-balancing circuit centered on MoOsm1 kinase governs adaptive responses to host-derived ROS in Magnaporthe oryzae. Elife 9:N/A (2020). PubMed: 33275098