Anti-SHP1 antibody [Y476] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

This IHC data was generated using the same anti-SHP1 antibody clone [Y476] in a different buffer formulation (cat# ab32559).

Unpurified ab32559, at a 1/50 dilution, staining human lymph node by immunohistochemistry, Paraffin embedded tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32559).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32559).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32559).

• WB

Lab

Western blot - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

This data was developed using the same antibody clone in a different buffer formulation (ab32559).

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.

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (<a href='/ko/products/primary-antibodies/shp1-antibody-y476-ab32559'>ab32559</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

PTPN6 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN6 knockout THP-1 cell line (<a href='/ko/products/cell-lines/human-ptpn6-knockout-thp-1-cell-line-ab273717'>ab273717</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 68 kDa,83 kDa

Observed band size: 70 kDa,85 kDa

false

• WB

Lab

Western blot - Anti-SHP1 antibody [Y476] - BSA and Azide free (AB226008)

This data was developed using ab32559, the same antibody clone in a different buffer formulation.

Western blot : Anti-SHP1 antibody [Y476] ab32559 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 68 kDa in Wild-type MCF7 cell lysates with a band observed at 55 kDa in PTPN6 CRISPR-Cas9 Edited MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (<a href='/ko/products/primary-antibodies/shp1-antibody-y476-ab32559'>ab32559</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 at 20 µg

Lane 2:

Western blot - Human PTPN6 knockout MCF7 cell line (ab287732) at 20 µg

Lane 3:

THP-1 at 20 µg

Lane 4:

SH-SY5Y at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa

false