Anti-Cytochrome C 抗体 [EPR1327]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunofluorescent analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Cytochrome C with unpurified ab133504 at 1/100 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunofluorescence staining of SH-SY5Y cells with purified ab133504 at a working dilution of 1 in 100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti rabbit (ab150077), used at a dilution of 1 in 500. ab7291 was used to stain tubulin, and this is shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom middle and right hand panels - for the negative controls, purified ab133504 was used at a dilution of 1/200 followed by an Alexa Fluor^® 594 goat anti-mouse antibody at a dilution of 1/500.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling Cytochrome C with ab133504 at a concentration of 0.1µg/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. ab133504 Anti-Cytochrome C antibody [EPR1327] 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling Cytochrome C with ab133504 at a concentration of 0.1µg/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. ab133504 Anti-Cytochrome C antibody [EPR1327] 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

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling Cytochrome C with unpurified ab133504 at 1/250 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-Cytochrome C antibody [EPR1327] (AB133504)

ab133504 (purified) at 1/30 immunoprecipitating Cytochrome C in Molt-4 cells.

For western blotting, a HRP-conjugated goat anti-rabbit (H+L), was used as the secondary antibody (1/1000).

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

All lanes:

Immunoprecipitation - Anti-Cytochrome C antibody [EPR1327] (ab133504)

Predicted band size: 11 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Immunohistochemical staining of paraffin embedded rat kidney with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Lab

Western blot - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution

All lanes:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution

All lanes:

Human fetal heart tissue lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution

All lanes:

Rat brain tissue lysate at 1/1000 dilution

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome C antibody [EPR1327] (AB133504)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/50000 dilution

All lanes:

Mouse brain tissue lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

• WB

Unknown

Western blot - Anti-Cytochrome C antibody [EPR1327] (AB133504)

All lanes:

Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/10000 dilution

Lane 1:

Molt4 lysate at 10 µg

Lane 2:

SH-SY5Y lysate at 10 µg

Lane 3:

Human heart lysate at 10 µg

Lane 4:

Human kidney lysate at 10 µg

Lane 5:

Human spleen lysate at 10 µg

Secondary

All lanes:

Goat-anti-rabbit HRP at 1/2000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Cytochrome C antibody [EPR1327] (AB133504)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.