Recombinant Anti-SHP1 antibody [Y476] (ab32559)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y476] to SHP1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SHP1 antibody [Y476]
See all SHP1 primary antibodies -
Description
Rabbit monoclonal [Y476] to SHP1 -
Host species
Rabbit -
Specificity
The antibody is predicted to detect isoforms 1, 2 and 3 of human SHP1 based on sequence analysis. -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human SHP1 aa 550 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P29350 -
Positive control
- WB: THP-1 cell lysate, A431 cell lysate, Jurkat cell lysate, K562 cell lysate. IHC-P: Human tonsil and lymph node tissue; Rat spleen tissue; Mouse liver tissue.
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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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y476 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32559 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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WB | (2) |
1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 68 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 68 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Plays a key role in hematopoiesis. This PTPase activity may directly link growth factor receptors and other signaling proteins through protein-tyrosine phosphorylation. The SH2 regions may interact with other cellular components to modulate its own phosphatase activity against interacting substrates. Together with MTUS1, induces UBE2V2 expression upon angiotensin II stimulation. -
Tissue specificity
Isoform 1 is expressed in hematopoietic cells. Isoform 2 is expressed in non-hematopoietic cells. -
Sequence similarities
Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.
Contains 2 SH2 domains.
Contains 1 tyrosine-protein phosphatase domain. -
Post-translational
modificationsPhosphorylated on serine and tyrosine residues. -
Cellular localization
Cytoplasm. Nucleus. In neurons, translocates into the nucleus after treatment with angiotensin II. - Information by UniProt
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Database links
- Entrez Gene: 5777 Human
- Entrez Gene: 15170 Mouse
- Entrez Gene: 116689 Rat
- Omim: 176883 Human
- SwissProt: P29350 Human
- SwissProt: P29351 Mouse
- SwissProt: P81718 Rat
- Unigene: 63489 Human
see all -
Alternative names
- 70Z-SHP antibody
- EC 3.1.3.48 antibody
- HCP antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)Immunohistochemical staining of paraffin embedded human tonsil with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : PTPN6 knockout THP-1 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab32559 observed at 70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab32559 was shown to react with SHP1 in wild-type THP-1 cells in Western blot with loss of signal observed in PTPN6 knockout sample. Wild-type THP-1 and PTPN6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab32559 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)Immunohistochemical staining of paraffin embedded rat spleen with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution (purified)
Lane 1 : SP2/0 cell lysate
Lane 2 : mouse marrow
Lane 3 : rat brain
Lane 4 : C6 cell lysate
Lane 5 : rat cerebral cortex
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 68 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution (purified)
Lane 1 : A431 cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 68 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)Immunohistochemical staining of paraffin embedded mouse liver with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution (unpurified)
Lane 1 : A- A431 cell lysate
Lane 2 : B- Jurkat cell lysate
Lane 3 : C- K562 cell lysate
Predicted band size: 68 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)
Unpurified ab32559, at a 1/50 dilution, staining human lymph node by immunohistochemistry, Paraffin embedded tissue.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)
Tissue Microarrays stained for "Anti-SHP1 antibody [Y476]” using "ab32559" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab32559 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (ab32559)
Tissue Microarrays stained for "Anti-SHP1 antibody [Y476]” using "ab32559" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab32559 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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
References (22)
ab32559 has been referenced in 22 publications.
- Dong Z et al. Hepatic Reduction in Cholesterol 25-Hydroxylase Aggravates Diet-induced Steatosis. Cell Mol Gastroenterol Hepatol 13:1161-1179 (2022). PubMed: 34990887
- Tathe P et al. SHP-1 dephosphorylates histone H2B to facilitate its ubiquitination during transcription. EMBO J 41:e109720 (2022). PubMed: 35938192
- Zhuang X et al. SHP-1 knockdown suppresses mitochondrial biogenesis and aggravates mitochondria-dependent apoptosis induced by all trans retinal through the STING/AMPK pathways. Mol Med 28:125 (2022). PubMed: 36273174
- 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
- Bian Y et al. SMURF1-mediated ubiquitylation of SHP-1 promotes cell proliferation and invasion of endometrial stromal cells in endometriosis. Ann Transl Med 9:362 (2021). PubMed: 33842583