Anti-Semaphorin 7a antibody [EPR22644-79]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Negative control image : IHC image of Semaphorin 7a staining in a section of human normal frozen liver* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab225602, 1ugml, 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. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Semaphorin 7a with ab255602 at 1/100 dilution (6.46μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human placenta. The section was incubated with ab255602 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• Flow Cyt

Unknown

Flow Cytometry - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Flow cytometric analysis of human peripheral blood mononuclear cells (PBMC) treated with 10μg/ml Phytohemagglutinin (PHA) for 2 days (Right pane;) / Untreated control (Left panel) labeling Semaphorin 7a with ab255602 at a 1/600 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150097) secondary antibody was used at a 1/5000 dilution. Gated on viable cells.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Semaphorin 7a with ab255602 at 1/100 dilution (6.46 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil. The section was incubated with ab255602 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

IHC image of Semaphorin 7a staining in a section of human normal frozen placenta* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab255602, 1ugml, 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. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre 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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

IHC image of Semaphorin 7a staining in a section of human normal frozen spleen* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab255602, 1ugml, 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. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre 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.

• IP

Unknown

Immunoprecipitation - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Semaphorin 7a was immunoprecipitated from 0.35 mg human testis lysate 10μg with ab255602 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab255602. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at a 1/1000 dilution.

Lane 1 : Human testis lysate 10μg
Lane 2 : ab255602 IP in Human testis lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab255602 in human testis lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-Semaphorin 7a antibody [EPR22644-79] (ab255602)

Predicted band size: 50 kDa,75 kDa

Observed band size: 75 kDa

false

• WB

Unknown

Western blot - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 26752048, 20962327).

All lanes:

Western blot - Anti-Semaphorin 7a antibody [EPR22644-79] (ab255602) at 1/1000 dilution

Lane 1:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

A549 treated with 100ng/ml TNF-alpha for 8 hours 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 1/100000 dilution

Predicted band size: 75 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-Semaphorin 7a antibody [EPR22644-79] (AB255602)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 26752048, 20962327).

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 10 secs; Lanes 2-3 : 37 secs.

All lanes:

Western blot - Anti-Semaphorin 7a antibody [EPR22644-79] (ab255602) at 1/1000 dilution

Lane 1:

Human tonsil lysate at 20 µg

Lane 2:

Human testis lysate at 20 µg

Lane 3:

Human placenta lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 75 kDa

false