Anti-Sall4 antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sall4 antibody (AB29112)

IHC image of Sall4 staining in a section of formalin-fixed paraffin-embedded human testis seminoma☆ performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab29112, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

☆Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

AbReview31910****

Immunocytochemistry/ Immunofluorescence - Anti-Sall4 antibody (AB29112)

ICC/IF image of ab29112 stained human embryonic stem cells. The cells were fixed in Paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 10% Goat serum, 0.1% BSA in PBS for 1 hour at RT, before incubation with ab29112 at a 1/100 dilution for 12 hours at 4°C. The secondary used was an Alexa Fluor^® 488 conjugated goat anti-rabbit polyclonal, at 1/200 dilution.

This image is courtesy of an anonymous abreview.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sall4 antibody (AB29112)

IHC image of Sall4 staining in a section of formalin-fixed paraffin-embedded human normal testis☆ performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab29112, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

☆Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sall4 antibody (AB29112)

IHC image of Sall4 staining in a section of formalin-fixed paraffin-embedded mouse normal testis performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab29112, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Sall4 antibody (AB29112)

ab29112 staining SALL4 in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab29112 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sall4 antibody (AB29112)

Negative control image : IHC image of Sall4 staining in a section of formalin-fixed paraffin-embedded mouse normal skeletal muscle performed on a Leica BOND™ system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab29112, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

• WB

Lab

Western blot - Anti-Sall4 antibody (AB29112)

Gel type : MOPS

Blocking buffer : 1% milk block

All lanes:

Western blot - Anti-Sall4 antibody (ab29112) at 1 µg/mL

Lane 1:

MEF1 whole cell lysate at 10 µg

Lane 2:

Mouse embryonic stem cell lysate at 10 µg

Lane 3:

HuES7 whole cell lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 112 kDa

Observed band size: 140 kDa

false

Exposure time: 12min

• WB

Project1912****

Western blot - Anti-Sall4 antibody (AB29112)

This antibody detects two bands which we believe correspond to the two isoforms of Sall4 outlined in Ma et al, 2006 (PMID : 16763212). This paper describes human isoforms running at 165 kDa and 95 kDa. In mouse embryonic stem cells we see both isoforms running at slightly lower molecular weights. We believe that these bands represent Sall4a (Swissprot : Q8BX22) and Sall4b (Swissprot : Q6S7E9) which are predicted to be 113 kDa and 66 kDa respectively.

All lanes:

Western blot - Anti-Sall4 antibody (ab29112) at 1 µg

All lanes:

E14tG2a (Mouse embryonic stem cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Predicted band size: 112 kDa

Observed band size: 140 kDa,85 kDa

false

• WB

Ap11662****

Western blot - Anti-Sall4 antibody (AB29112)

MEF1 whole cell lysate was included as a negative control.

Secondary antibody - goat anti-rabbit HRP (ab97051)

All lanes:

Western blot - Anti-Sall4 antibody (ab29112) at 1 µg/mL

Lane 1:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 2:

E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 112 kDa

Observed band size: 140 kDa,60 kDa,80 kDa

true

Exposure time: 8min

• IHC-P

AbReview30908****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sall4 antibody (AB29112)

IHC-P image of Sall4 staining with ab29112 on tissue sections from a 3 year old adult marmoset testis. The sections were subjected to heat-mediated antigen retrieval using Dako antigen retrieval solution. The sections were then blocked with 5% milk for 30 minutes at 25°C, before incubation with ab29112 (1/100 dilution) for 20 hours at 4°C. The secondary was an Alexa Fluor^® 555 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

This image is courtesy of an abreview by Zachary Yu-Ching Lin.

• WB

Lab

Western blot - Anti-Sall4 antibody (AB29112)

Western blot : Anti-SALL4 antibody (ab29112) staining at 1 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab29112 was shown to bind specifically to SALL4. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Sall4 antibody (ab29112) at 1 µg/mL

Lane 1:

MEF cell lysate at 20 µg

Lane 2:

mESC cell lysate at 20 µg

Lane 3:

HuES7 cell lysate at 20 µg

Lane 4:

Mouse Testis cell lysate at 20 µg

Lane 5:

Human testis cell lysate at 20 µg

Lane 6:

MCF7 cell lysate at 20 µg

Lane 7:

NCCIT cell lysate at 20 µg

Lane 8:

HeLa cell lysate at 20 µg

Lane 9:

Empty cell lysate

Lane 10:

Recombinant Human Sall4 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sall4-protein-ab153444'>ab153444</a>) cell lysate at 0.5 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 140 kDa

false