Anti-Thrombospondin 1 antibody [EPR22928-10]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

Immunohistochemical analysis of paraffin-embedded human bone marrow tissue labeling Thrombospondin 1 with ab263905 at 1/5000 dilution 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 ab263905 for 15 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263905).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Thrombospondin 1 with ab263905 at 1/5000 dilution 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 ab263905 for 15 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263905).

• WB

Lab

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

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

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 8422970, 27588705).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (ab263905) 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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/25000 dilution

Predicted band size: 129 kDa

Observed band size: 180 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

Thrombospondin 1 was immunoprecipitated from 0.35 mg Mouse platelet lysate 10ug with ab263905 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263905 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : Mouse platelet lysate 10ug

Lane 2 : ab263905 IP in Mouse platelet lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab263905 in Mouse platelet lysate

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

Exposure time : 3 min

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263905).

All lanes:

Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22928-10] (ab263905)

Predicted band size: 129 kDa

false

• WB

Supplier Data

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

Exposure time : 1 second.

Observed MW : 180 kDa.

All lanes:

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (ab263905) at 1/1000 dilution

All lanes:

Mouse platelet lysate at 20 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 1/25000 dilution

Predicted band size: 129 kDa

false

• WB

Lab

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (AB263905)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1 : 59 seconds Lane 2 : 15 seconds The identity of the lower MW bands at approximately 25 kDa is unknown. The bands beneath the target band (~75-180 kDa) are likely to be degraded target fragments.

Lanes 1 - 2:

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/thrombospondin-1-antibody-epr22928-10-bsa-and-azide-free-ab263952'>ab263952</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-Thrombospondin 1 antibody [EPR22928-10] (ab263905) at 1/1000 dilution

Lane 1:

Mouse platelet whole cell lysate at 20 µg

Lane 2:

Rat platelet whole cell lysate at 20 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: 129 kDa

Observed band size: 180 kDa

false