Product datasheet
Anti-SHP1 antibody ab2020
2 Abreviews 5 References 6 Images
Overview
Product name
Anti-SHP1 antibody
Description
Rabbit polyclonal to SHP1
Specificity
Recognises the recombinant phosphatase domain fusion protein (60 kDa) in Western blots.
Predicted molecular weight for SHP1 is 70 kDa. When used in IHC staining on human tonsil
stains the mantle zone of B cell follicles and T cells in the interfollicular area. Seems to be
selective for small lymphoid cells, which is in keeping with published data eg. Khoury J et al. and
Oka T et al.
Tested applications
IHC-P, IP, WB, ICC/IF
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human
SHP1.Read Abcam's proprietary immunogen policy(Peptide available as ab82208.)
Positive control
This antibody gave a positive signal in SHP1 Recombinant protein and Jurkat Whole Cell
Lysate.
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 or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab2020 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
IHC-P
Abreviews
Notes
Use at an assay dependent concentration.
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Application
Abreviews
Notes
IP
Use a concentration of 5 µg/ml.
WB
1/500 - 1/1000. Detects a band of approximately 68 kDa (predicted molecular
weight: 60 kDa).
ICC/IF
Use a concentration of 1 µg/ml.
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
modifications
Phosphorylated on serine and tyrosine residues.
Cellular localization
Cytoplasm. Nucleus. In neurons, translocates into the nucleus after treatment with angiotensin II.
Anti-SHP1 antibody images
ab2020 was used in immunohistochemistry
with paraffin embedded sections of human
tonsil, using DAB as a chromogen (brown).
Counterstaining of nuclei was performed with
haemotoxylin (blue).
Picture of a B cell follicle.
The antibody shows strong staining of mantle
zone cells and some staining of a few cells in
- SHP1 antibody (ab2020)
the light zone of the germinal centre. This is in
keeping with published data (see Khoury J et
al and Oka T et al.).
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ICC/IF image of ab2020 stained human HeLa
cells. The cells were PFA fixed (10 min),
permabilised in TBS-T (20 min) and
incubated with the antibody (ab2020, 1µg/ml)
for 1h at room temperature. 1%BSA / 10%
normal goat serum / 0.3M glycine was used to
quench autofluorescence and block nonspecific protein-protein interactions. The
Immunocytochemistry/ Immunofluorescence -
secondary antibody (green) was Alexa Fluor®
SHP1 antibody (ab2020)
488 goat anti-rabbit IgG (H+L) used at a
1/1000 dilution for 1h. Alexa Fluor® 594 WGA
was used to label plasma membranes (red).
DAPI was used to stain the cell nuclei (blue).
All lanes : Anti-SHP1 antibody (ab2020) at 1
µg/ml
Lane 1 : PTPN6 - Recombinant Protein
(Human) at 0.1 µg
Lane 2 : Jurkat (Human T cell lymphoblastlike cell line) Whole Cell Lysate at 10 µg
Lane 3 : A431 (Human epithelial carcinoma
cell line) Whole Cell Lysate at 10 µg
Lane 4 : K562 (Human erythromyeloblastoid
leukemia cell line) Whole Cell Lysate at 10 µg
Western blot - SHP1 antibody (ab2020)
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 60 kDa
Observed band size : 68,92 kDa
Additional bands at : 45 kDa. We are
unsure as to the identity of these extra bands.
Exposure time : 30 seconds
3
SHP1 was immunoprecipitated using 0.5mg
Jurkat whole cell extract, 5µg of Rabbit
polyclonal to SHP1 and 50µl of protein G
magnetic beads (+). No antibody was added
to the control (-).
The antibody was incubated under agitation
with Protein G beads for 10min, Jurkat whole
cell extract lysate diluted in RIPA buffer was
Immunoprecipitation - Anti-SHP1 antibody
(ab2020)
added to each sample and incubated for a
further 10min under agitation.
Proteins were eluted by addition of 40µl SDS
loading buffer and incubated for 10min at
70oC; 10µl of each sample was separated on
a SDS PAGE gel, transferred to a
nitrocellulose membrane, blocked with 5%
BSA and probed with ab2020.
Secondary: Mouse monoclonal [SB62a]
Secondary Antibody to Rabbit IgG light chain
(HRP) (ab99697).
Band: 68kDa; SHP1.
All lanes : Anti-SHP1 antibody (ab2020) at
1/2000 dilution
Lane 1 : PTP1B (PTPN1)
Lane 2 : TCPTP (PTPN2)
Lane 3 : PTPH1 (PTPN3)
Western blot - SHP1 antibody (ab2020)
Lane 4 : MEG1 (PTPN4)
Lane 5 : STEP (PTPN5)
Lane 6 : SHP1 (PTPN6)
Lane 7 : HePTP (PTPN7)
Lane 8 : SHP2 (PTPN11)
Lane 9 : BDP1 (PTPN18)
Predicted band size : 60 kDa
Observed band size : 60 kDa
Recombinant PTP phosphatase domains
were probed with ab2020 at 1/2000. The
antibody recognises the recombinant SHP1
fusion protein at ~60 kDa.
Data provided by Purely Proteins
4
ab2020 was used in immunohistochemistry
with paraffin embedded sections of human
tonsil, using DAB as a chromogen (brown).
Counterstaining of nuclei was performed with
haemotoxylin (blue).
The antibody shows staining of T cells in the
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - SHP1 antibody
(ab2020)
interfollicullar area. This is in keeping with
published data (see Khoury J et al and Oka T
et al.).
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