Anti-Thrombospondin 1 抗体 [EPR22927-54]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labelling Thrombospondin 1 with ab267388 at 1/5000 (0.101 μg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a Ready to use dilution. Positive staining on extracellular matrix of human cervical carcinoma. The section was incubated with ab267388 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunohistochemical analysis of paraffin-embedded human bone marrow tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the megakaryocytes in the human bone marrow (PMID : 28239144). The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling Thrombospondin 1 with ab267388 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the platelets in the human spleen (PMID : 28239144). The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab267388).

Thrombospondin 1 was immunoprecipitated from 0.35 mg HUVEC (Human umbilical vein endothelial cell) whole cell lysate 10ug with ab267388 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267388 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate 10ug
Lane 2 : ab267388 IP in HUVEC whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267388 in HUVEC whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)

Predicted band size: 129 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunofluorescent analysis of 100% methanol-fixed, permeabilized HUVEC (human umbilical vein endothelial cell) cells labeling Thrombospondin 1 with ab267388 at 1/100 dilution (5 µg/ml), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution. 100% methanol fixation is recommended.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 3T3-L1 (mouse embryonic fibroblast) starved with 0.4% serum for 24h, then cultured with 15% serum for 6h (Red) / Untreated control (Green) cells labelling Thrombospondin 1 with ab267388 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Intracellular Flow Cytometry analysis of PC-12 (Rat adrenal gland pheochromocytoma cell line) cells labeling Thrombospondin 1 with ab267388 at 1/500 dilution (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat Anti-rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (Black) (ab172730). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the megakaryocytes and platelets in the mouse spleen is observed. The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Immunofluorescent analysis of 100% Methanol-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Thrombospondin 1 with ab267388 at 1/100 (5.3 μg/ml) dilution, followed by ab150077 AlexaFluor_® 488 Goat anti-Rabbit secondary antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor_® 594) was used to counterstain tubulin at 1/200 (2.5 μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor_® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• IP

Unknown

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab267388).

Thrombospondin 1 was immunoprecipitated from 0.35 mg Mouse platelets whole cell lysate 10ug with ab267388 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267388 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse platelets whole cell lysate 10ug
Lane 2 : ab267388 IP in Mouse platelets whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267388 in Mouse platelets whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)

Predicted band size: 129 kDa

false

• WB

Supplier Data

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

The full-length TSP 1 (180kDa) and a 160-kDa band, likely to be an TSP 1 isoform or fragment, are observed.

The molecular weight observed is consistent with what has been described in the literature (PMID : 1426766, 27588705).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

Mouse platelet lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 129 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

TSP 1 is a serum-responsive gene. Its express is elevated in quiescent 3T3 with serum addition.

The full-length TSP 1 (180kDa) and a 160-kDa band, likely to be an TSP 1 isoform or fragment, are observed.

The molecular weight observed is consistent with what has been described in the literature (PMID : 1426766, 27588705).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution

Lane 1:

3T3-L1 (mouse embryonic fibroblast) starved with 0.4% serum for 30 hours whole cell lysate at 10 µg

Lane 2:

3T3-L1 starved with 0.4% serum for 24 hours, then cultivated with 15% serum for 6 hours. whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 129 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (AB267388)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST The full-length TSP 1 (180 kDa) and a ~140 kDa band, likely to be a TSP 1 isoform or fragment, are observed. The molecular weight observed is consistent with what has been described in the literature (PMID : 1426766, 27588705).

Lane 1:

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (<a href='/products/primary-antibodies/thrombospondin-1-antibody-epr22927-54-bsa-and-azide-free-ab267397'>ab267397</a>) at 1/1000 dilution

Lane 1:

Western blot - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution

All lanes:

Rat platelet whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 129 kDa

Observed band size: 140 kDa,180 kDa

false

Exposure time: 5.5s