MA5-11674

antibody from Invitrogen Antibodies

Targeting: CYCS CYC, HCS

Provider product page for MA5-11674

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [41]
Comments [0]
Validations
Immunocytochemistry [1]
Immunohistochemistry [4]
Other assay [5]

Submit

Product number: MA5-11674 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cytochrome C Monoclonal Antibody (7H8.2C12)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA5-11674 targets Cytochrome c in IHC (P) and WB applications and shows reactivity with Human, mouse, and Rat samples. The MA5-11674 immunogen is synthetic peptides corresponding to amino acids 1-80, 81-104 and 66-104 of pigeon cytochrome c.
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: 7H8.2C12
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

CYCS protein structure - MA5-11674 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human heart stained with cytochrome c antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of cardiac muscle.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytochrome c showing staining in the cytoplasm of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a using Cytochrome c Mouse Monoclonal Antibody (Product # MA5-11674) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytochrome c showing staining in the cytoplasm of paraffin-embedded human colon carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cytochrome c Mouse Monoclonal Antibody (Product # MA5-11674) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytochrome c showing staining in the cytoplasm of paraffin-embedded mouse skeletal muscle tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cytochrome c Mouse Monoclonal Antibody (Product # MA5-11674) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Esculetin induces loss of Mitochondrial membrane potential: a Flow cytometric analysis of PANC-1 cells stained with JC-1 dye after 100 muM esculetin treatment for indicated time showed a temporal decrease in ratio of red fluorescence (JC-1 aggregates) to green florescence (JC-1 monomers). b Percentage of cells exhibiting monomers and aggregates of JC-1 after treatment of cells with esculetin for different time intervals. c Western blot analysis showing temporal increase in cytosolic cytochrome C in PANC-1 cells. (VC stands for vehicle control, E stands for esculetin treatment sample for indicated time, CCCP stands for positive control i.e., carbonyl cyanide 3-chlorophenylhydrazone treated cells, numerals represent time of esculetin treatment). Data represents the mean +- SD of three independent experiments. The significance was determined using ANOVA (Bonferroni's test). Key:* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Simvastatin attenuated cytochrome C expression in renal tubular epithelial cells after LPS-administration. The expression of cytochrome C in rat renal tissue examined by immunohistochemical staining, magnification 400x, ( A ) negative immunostaining in the control group, ( B ) intense brown cytoplasmic staining in the tubular epithelium in the LPS group indicates increased expression of the pro-apoptotic protein cytochrome C. A significant decrease of cytochrome C-positive cells in the groups pretreated with simvastatin 20 mg/kg ( C ) and 40 mg/kg group ( D ), respectively. A quantitative analysis of cytochrome C-positive renal tubular epithelial cells assessed in the immunohistochemically stained sections in the renal tissue (* p < 0.05 vs. LPS group, ** p < 0.01 vs. simvastatin 20 group, # p < 0.01 vs. LPS group) ( E ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Western blot analysis: The cell signaling protein expressions of ( a ) BT-474 and ( b ) MDA-MB-231 cells using PBS, PTX alone, TTZ alone, and PTXNR-TTX treatments. The relative intensity of XIAP, actin, cleaved caspase-9, caspase 3, cleaved caspase 3, and cytochrome C is analyzed after 48 h of treatment in ( c ) BT-474 and ( d ) MDA-MB-231 cells using ImageJ/ Fiji. For each protein expression the experiment was replicated at least n = 2-4 times and the average data is presented as mean +- standard deviation. The relative protein expressions are normalized to the untreated control cells and to the housekeeping GAPDH protein expression (hypothesized expression mean, u = 1). The p values for overexpression or downregulation of cytochrome-C, cleaved caspase-3, and XIAP in BT-474 are 0.039, 0.003, and 0.000049, respectively. The p value for the overexpression of cytochrome-C in MDA-MB-231 is 0.047. ***Represents p < 0.05 (for detailed p value calculations, please see SI Table 6 ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Simvastatin attenuated cytochrome C expression in alveolar epithelial cells in LPS-induced inflammation. Representative images of pulmonary parenchyma that were challenged with LPS or pretreated with 20 mg/kg of simvastatin or 40 mg/kg of simvastatin, respectively, prior to LPS, assessed by immunohistochemistry (400x). ( a ) Control group. ( b ) Intense cytoplasmic staining of cytochrome C (arrows) in alveolar epithelium, macrophages, and endothelial cells in the LPS group. Marked reduction of cytochrome C-positive cells in the groups pretreated with 20 mg/kg of simvastatin ( c ) and 40 mg/kg of simvastatin prior to LPS ( d ). Semiquantitative analysis of cytochrome C-positive cells in pulmonary parenchyma assessed in the immunohistochemically stained sections in the lung tissue (* p < 0.05 vs. LPS group, ** p < 0.01 vs. LPS group, # p < 0.05 vs. 20 mg/kg of simvastatin) ( e ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 miR-26a/b target PKCdelta to inhibit apoptosis in oral keratinocytes. a - d Western blot showing changes of cleaved caspase 9, cleaved PARP, and cleaved caspase 3 levels in HOKs added with miR-26a/b mimics ( a ) or inhibitors ( b ). c - d Western blot analysis of cleaved caspase 9, cleaved PARP, and cleaved caspase 3 expression in mouse oral keratinocytes. C57BL/6 mice were subject to miR-26a/b mimics ( c ) or inhibitors ( d ) by tail vein injection. e Putative miR-26a/b binding site in the 3'UTR of PKCdelta mRNA. f Schematic illustration of binding site location in the 3'UTR of hPKCdelta cDNA. g Luciferase reporter assay showing miR-26a/b target binding site in the 3'UTR of hPKCdelta in HOKs which were transfected with Luc-PKCdelta-3'UTR or pRL-TK (control) plasmids. Luciferase activity was quantified after 24 h. h HOKs were co-transfected with Luc-PKCdelta-3'UTR or Luc-PKCdelta-3'UTR-Mut and miR-26a/b inhibitors as indicated. i-j PKCdelta expression and phosphorylation in HOKs with miR-26a/b mimics or inhibitors were measured by real-time PCR ( i ) or western blot ( j ). Western blot showing Bax translocation, cyt c release, and downstream apoptotic factors expression in HOKs during miR-26a/b mimics ( k ) or inhibitors ( l ) treatment with or without PKCdelta plasmids or siRNA transfection as indicated. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. corresponding control; n = 3 for assays in vitro and n = 5 for assays in vivo. Ctrl control, Mito mitochondria, Cyto cytopl