Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

This data was developed using the same antibody clone in a different buffer formulation (ab200999).

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling C3/C3b with ab200999 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

Anti-C3/C3b antibody [EPR19394] ab200999 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

The plasma in blood vessels of rat kidney was positive staining. The staining pattern is similar to what has been observed in the literature (PMID : 23104558).

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab200999).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on hepatocytes of rat liver is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab200999).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

The plasma in blood vessel of mouse cardiac muscle was positive staining. The staining pattern is similar to what has been observed in the literature (PMID : 23104558).

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab200999).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

The plasma in blood vessels of rat lung was positive staining. The staining pattern is similar to what has been observed in the literature (PMID : 23104558).

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab200999).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

The plasma in blood vessels of mouse cerebral cortex was positive staining.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab200999).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

This IHC data was generated using the same anti-C3 antibody clone [EPR19394] in a different buffer formulation (cat# ab200099).

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling C3 with ab200999 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on hepatocytes of mouse liver is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-C3 antibody [EPR19394] - Low endotoxin, Azide free (AB225539)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200999).

Western blot : Rabbit Monoclonal [EPR19394] to C3/C3b ab200999 staining at 1/2000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 170 kDa in Wild-type 3T3-L1 after treatment with brefeldin A, with no signal observed at this size in C3 knockout 3T3-L1 samples. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-C3 antibody [EPR19394] (<a href='/ko/products/primary-antibodies/c3-antibody-epr19394-ab200999'>ab200999</a>) at 1/2000 dilution

Lane 1:

Wild-type 3T3-L1 Treated: Brefeldin A (300 ng/mL, 24 h) cell lysates at 20 µg

Lane 2:

Wild-type 3T3-L1 Vehicle Control for Brefeldin A (0 ng/mL, 24 h) cell lysates at 20 µg

Lane 3:

C3 knockout 3T3-L1: Treated: Brefeldin A (300 ng/mL, 24 h) cell lysates at 20 µg

Lane 4:

C3 knockout 3T3-L1 Vehicle Control for Brefeldin A (0 ng/mL, 24 h) cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 187 kDa

Observed band size: 170 kDa

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