MM700B

antibody from Invitrogen Antibodies

Targeting: IFNG

Provider product page for MM700B

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [84]
Comments [0]
Validations
Western blot [3]
Other assay [4]

Submit

Product number: MM700B - Provider product page
Provider: Invitrogen Antibodies
Product name: IFN gamma Monoclonal Antibody (XMG1.2), Biotin
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MM700B targets IFN gamma in ELISA, IHC-F, Neutralization and WB applications and shows reactivity with mouse and human samples.
Conjugate: Biotin
Antibody clone number: XMG1.2
Concentration: 0.5 mg/mL

IFNG protein structure - MM700B shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of IFN Gamma was performed by loading 20 µg of indicated whole cell lysates in reducing sample buffer and 8 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IFN Gamma was detected at ~20 kDa using a IFN Gamma rat monoclonal biotin conjugate antibody MM700B at a concentration of 2 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Streptavidin-HRP conjugate (Product # 21126) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of IFN Gamma was performed by loading indicated amount of Human IFN Gamma recombinant protein in reducing sample buffer and 8 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IFN Gamma was detected at ~16 kDa using a IFN Gamma rat monoclonal biotin conjugate antibody MM700B at a concentration of 2 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Streptavidin-HRP conjugate (Product # 21126) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of IFN Gamma was performed by loading indicated amount of Human IFN Gamma recombinant protein in reducing sample buffer and 8 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IFN Gamma was detected at ~16 kDa using a IFN Gamma rat monoclonal biotin conjugate antibody MM700B at a concentration of 2 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Streptavidin-HRP conjugate (Product # 21126) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Direct ELISA analysis of IFN gamma was performed by coating wells of a 96-well plate with 100 µL per well of Human IFN gamma (Product # RIFNG100) diluted in carbonate/bicarbonate buffer (Product # 28382), starting at a concentration of 1000pg/mL and serially diluting 2-fold to a concentration of 1 µg/mL, overnight at 4°C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse anti-IFN gamma Biotin conjugated monoclonal antibody (Product # MM700B) at a concentration of 1 µg/mL for 1 hour at 37C. The plate was washed, then incubated with 100 µL per well of an streptavidin HRP-conjugate (Product # 21126) at a dilution of 1:5000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 10 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550 nm.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA of IFN gamma was performed by coating wells of a 96-well plate with 100 µL of an IFN gamma mouse monoclonal antibody (Product # MM700) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4°C. Wells were blocked with 200 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 28382) for 2 hours, and 100 µL of recombinant mouse IFN gamma protein ranging from 16-3000 pg/mL were incubated for 1 hour at room temperature. The plate was washed with 1X PBS (Product # 28348), and 100 µL per well of an IFN gamma Biotin conjugated mouse monoclonal antibody (Product # MM700B) diluted to a concentration of 1 µg/mL was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a Streptavidin-HRP (Product # 21126) was incubated for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028) for 15-30 minutes, followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Corylin improves the TLR4 signal pathway in DSS-induced colitis mice. ( A ) Expression analysis by immunohistochemical staining of TLR4, p-p38, and AP-1 in mice after the indicated treatment. Scale bar: 100 mum. Red arrows highlight positive staining. ( B ) Western blot analysis of TLR4, MyD88, p-p38, AP-1, and beta-actin (loading control) in colon homogenates. Right graph indicates quantification relative to beta-actin. ( C , D ) mRNA expression of Ifngamma , Tnf-alpha , Il-6 , Il-1beta , Nlrp3 , Asc , Pannexin , and Pro-caspase 1 determined by qRT-PCR. ( E , F ) Western blot analysis of IFNgamma, TNF-alpha, IL-6, IL-1beta, NLRP3, ASC, Pannexin, and Caspase 1, and beta-actin (loading control) in colon homogenates. Results represent mean +- SEM. * p < 0.05, Normal compared with DSS; # p < 0.05, DSS compared with DSS + Corylin (L); Y= p < 0.05, DSS compared with DSS + Corylin (H); SS p < 0.05, DSS + Corylin (L) compared with DSS + Corylin (H). DSS, dextran sodium sulfate; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation primary response 88; AP-1, activator protein 1; Ifngamma , interferon gamma; Tnf-alpha , tumor necrosis factor-alpha; Il-6 , interleukin-6; Nlrp3 , NACHT, LRR, and PYD domains-containing protein 3.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Corylin improves the TLR4 signaling pathway in AOM/DSS-induced-colitis-associated colorectal cancer mice. ( A ) Expression analysis by immunohistochemical staining of TLR4, p-p38, and AP-1 in mice after the indicated treatment. Scale bar: 100 mum. ( B ) Western blot analysis of TLR4, MyD88, p-p38, AP-1, and beta-actin (loading control) in colon. ( C , D ) mRNA expression of Ifngamma , Tnf-alpha , Il-6 , Il-1beta , Nlrp3 , Asc , Pannexin , and Pro-caspase 1 as determined by qRT-PCR. ( E , F ) Western blot analysis of IFNgamma, TNF-alpha, IL-6, IL-1beta, NLRP3, ASC, Pannexin, and Caspase 1, and beta-actin (loading control) in colon homogenates. Results represent mean +- SEM. * p < 0.05, Normal compared with AOM/DSS; # p < 0.05, AOM/DSS compared with AOM/DSS + Corylin (L); Y= p < 0.05, AOM/DSS compared with AOM/DSS + Corylin (H); SS p < 0.05, DSS + Corylin (L) compared with DSS + Corylin (H). AOM, azoxymethane; DSS, dextran sodium sulfate; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation primary response 88; AP-1, activator protein 1; Ifngamma , interferon gamma; Tnf-alpha , tumor necrosis factor-alpha; Il-6 , interleukin-6; Nlrp3 , NACHT, LRR, and PYD domains-containing protein 3.
Conjugate: Biotin