MA5-14568

antibody from Invitrogen Antibodies

Targeting: PTGS2 COX2

Provider product page for MA5-14568

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [61]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Immunohistochemistry [3]
Other assay [5]

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Product number: MA5-14568 - Provider product page
Provider: Invitrogen Antibodies
Product name: COX2 Monoclonal Antibody (SP21)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA5-14568 targets COX2 in IHC (P) applications and shows reactivity with Human samples.
Antibody clone number: SP21
Concentration: 0.006 mg/mL

PTGS2 protein structure - MA5-14568 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of COX2 showing staining in the cytoplasm and membrane 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 COX2 monoclonal antibody (Product # MA5-14568) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°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 COX2 showing staining in the cytoplasm and membrane 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 COX2 monoclonal antibody (Product # MA5-14568) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°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: Formalin-fixed, paraffin-embedded breast carcinoma stained with COX2 antibody using peroxidase conjugate and AEC chromogen. Note cytoplasmic staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 COX-2, phopho-IkappaBalpha, and reactive oxygen species expression in LPS-stimulated BV-2 cells with and without L-VGCC blockade. Effects of L-VGCC blockade on COX-2 expression ( a ) and detection of phosphorylated IkappaBalpha ( b ) in BV-2 cells stimulated with LPS. Quantitative RT-PCR analysis of COX-2 expression in BV-2 cells treated with DMSO control (2 mul/ml), Zinc20267861 (2.0 muM), Zinc18204217 (2.0 muM), and Zinc33254827 (2.5 muM) stimulated with LPS 1 mug/ml ( c ). Arg-1 expression with the same treatments in BV-2 cells stimulated with IL-4 (20 ng/ml) ( d ) ( n = 3 with 3 technical replicates). A one-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. p > 0.05; *** p < 0.001. Target genes were normalized to cyclophilin and presented as the fold change of DMSO control. Detection of reactive oxygen species in BV-2 cell culture, minimal oxidation of dihydroethidium in DMSO 2 mul/ml untreated control cells ( e ); strong oxidized ethidium signals detected when stimulated with LPS 1 mug/ml ( e ). Treatment with 3.5 muM of amlodipine ( g ), Zinc20267861 (2.0 muM) ( h ), Zinc18204217 (2.0 muM) ( i ), and Zinc33254827 (2.5 muM) ( j ) demonstrated marked reduction of the oxidized ethidium signals in BV-2 cell culture.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Effects of L-VGCC blockade on laser-induced CNV and expression of COX-2 and phospho-Ikappabalpha in mice. Fundus in vivo images of vehicle-treated laser CNV fundus 5 days after induction of the model ( a ) and Zinc20267861 (10 mug) subconjunctival treated eyes ( e ). Arrows indicate laser burn spots. In vivo fluorescent angiography in vehicle-treated laser CNV animals 30 ( b ), 60 ( c ), and 120 ( d ) after subcutaneous injection of fluorescein sodium 1 mg/kg and same time points ( f-h ) for Zinc20267861 (10 mug) subconjunctival treated eyes. Arrows indicate fluorescein leakage areas. Ex vivo evaluation of the laser spot sizes in retinal pigment epithelia choroid-scleral complexes (RCSC) flat mounts in control ( i ) and Zinc20267861 (10 mug) treated eyes ( j ) counterstained with GSA-lectin. Immunofluorescent detection of COX-2 ( k , l ) and phospho-Ikappabalpha ( m , n ) in RCSC flat mounts in control and Zinc20267861 - treated eyes. Quantitative analysis of fluorescein leakage area over time in vivo ( o ). A one-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. *** p < 0.001; ( n = 3). Ex vivo quantitative analysis of fluorescein-stained neovessels ( n = 6) ( p ), retinal pigment epithelia disruption area ( n = 6) ( q ), and GSA-lectin stained neovessels ( n = 6) ( r ). Student t -test was used to determine statistical significance between two groups. * p < 0.05; ** p < 0.01; *** p < 0.001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Expression level of protein of COX-2 and 5-LOX through western blot and levels of gene contributor through quantitative RT-PCR. Immunoblotting of respective individual group [I-Control (Normal saline, 3 ml/kg, p.o.), II- Toxic control (MNU 47 mg/kg, i.v.), III- Zaltoprofen (10 mg/kg, p.o. + MNU 47 mg/kg, i.v.), IV- Zileuton (10 mg/kg, p.o. + MNU 47 mg/kg, i.v.) and V- Zaltoprofen + Zileuton (5 + 5 mg/kg, p.o. + MNU 47 mg/kg, i.v.)] for COX-2 and 5-LOX. Excised mammary gland tissue sample lysed in trizol for RNA extraction and analyzed for the mRNA expression of COX-2 and 5-LOX by qRT-PCR. beta-actin was used as loading control. Each experiment was performed in triplicate. Values are presented as Mean +- SD. Comparisons were made by the one-way ANOVA followed by Bonferroni multiple test. All groups were compared to the MNU treated group ( ** p < 0.01, *** p < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 1 (A) Osteonectin, chondrosarcoma grade III. (B) Osteonectin, chondromyxoid fibroma. (C) Bcl-2, chondrosarcoma grade II. (D) Cox-2, chordoma. (E) Actin, chondroblastoma. (F) Calponin, the single chondromyxoid fibroma with a ++ reaction. (G) Mdm-2, chordoma. (H) D2-40 (podoplanin), advancing edge of chordoma with soft tissue invasion. The only positivity is seen in compressed lymph vessels. (I) D2-40 (podoplanin), chondrosarcoma grade II. (J) YKL-40, chondrosarcoma grade II. (K) YKL-40, ESMC. Note staining of the contents in the blood vessel. (L) CD117, ESMC. All magnifications originally x 200.