Antibodies, Proteins, Kits and Reagents for Life Science

345 091 ₸ Product size

100 µl 345 091 ₸ 10 µl 137 366 ₸

Order now and get it on Tuesday March 02, 2021

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

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 & Species

Application	Species
IHC-P	Mouse Rat Human
WB	Mouse Rat Human

See all applications and species data

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
For more information see here.
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.

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

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.
Western blot - Anti-SHP1 antibody [Y476] (ab32559)

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.
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.
Western blot - Anti-SHP1 antibody [Y476] (ab32559)

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
Western blot - Anti-SHP1 antibody [Y476] (ab32559)

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.
Western blot - Anti-SHP1 antibody [Y476] (ab32559)

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.
Anti-SHP1 antibody [Y476] (ab32559)

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