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
-
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.
-
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
-
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...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y476 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
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 |
---|---|---|
WB | (2) |
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.
|
Notes |
---|
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
-
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
-
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
-
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.
-
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.
-
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.
-
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 -
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.
-
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? -
Unpurified ab32559, at a 1/50 dilution, staining human lymph node by immunohistochemistry, Paraffin embedded tissue.
-
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.
-
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
-
SDS download
-
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