Mouse anti-Human Insulin Monoclonal Antibody | anti-INS antibody

Mouse Monoclonal [clone 2D11.H5] (IgG1,k) to Human Insulin

Cat. Num. AAA51775

• Gene Names: INS; IDDM; ILPR; IRDN; IDDM1; IDDM2; MODY10
• Reactivity: Human
• Applications: ELISA, Western Blot, Immunohistochemistry
• Purity: Protein A Purified
• Synonyms: Insulin, Antibody; Mouse Monoclonal [clone 2D11.H5] (IgG1,k) to Human Insulin; Anti-Insulin Antibody (clone 2D11.H5) IHC-plus; IRDN; IDDM2; Preproinsulin; ILPR; Insulin; MODY10; Proinsulin; Human Insulin; anti-INS antibody

• Host: Mouse
• Reactivity: Human
• Clonality: Monoclonal
• Isotype: IgG1,k
• Clone Number: 2D11.H5
• Specificity: Reacts specifically with insulin from human and swine sources. Cross reactivity with insulin from mouse and rat does not occur. Cross reactivity with insulin from other sources has not been determined.
• Purity/Purification: Protein A Purified
• Form/Format: 0.02 M Potassium Phosphate, pH 7.2, 0.15 M NaCl, 0.01% Sodium Azide
• Concentration: 1 mg/ml (varies by lot)
• Sequence Length: 110

• Applicable Applications for anti-INS antibody: ELISA, WB (Western Blot), IHC (Immunohistochemistry)
• Target Species: Insulin
• Family / Subfamily: Hormone / Not assigned-Hormone
• Immunogen Species: Human
• Immunogen: This protein A purified monoclonal antibody was produced by repeated immunizations with purified human insulin coupled to bovine serum albumin (BSA).
• Disclaimer: Due to the highly specific nature of antibodies and antigens, we cannot predict or be held responsible with respect to how this antibody will behave in your systems. Researchers using this antibody should conduct optimization studies to achieve the most optimal result possible for their intended application.
• Recommended Immunohistochemistry Protocol: The following protocol is a recommendation only, and AAA Biotech makes no guarantee of the results:; ; Tissue Preparation: ; Formalin fixation and embedding in paraffin wax. ; ; Tissue Sectioning: ; Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C. ; ; Deparaffinization: ; Wash dry slides in 3 changes of xylene - 5 minutes each @ RT; ; Rehydration: ; Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT; Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT; Wash slides in 1 change of 80% alcohol - 3 minutes @ RT; Rinse slides in gentle running distilled water - 5 minutes @ RT; ; Antigen retrieval: ; Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes; Remove from heat and let stand at room temperature in buffer - 20 minutes; Rinse in 1X TBS with Tween (TBST) -1 minute @ RT; ; Immunostaining: ; (Do not allow tissues to dry at any time during the staining procedure) ; Apply a universal protein block - 20 minutes @ RT; Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT; Rinse slides in 1 X TBST - 1 minute @ RT; Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT; Rinse slides in 1X TBST -1 minute @ RT; Apply alkaline phosphatase streptavidin - 30 minutes @ RT; Rinse slides in 1X TBST -1 minute @ RT; Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT; Wash slides in distilled water - 1 minute @ RT; ; Dehydrate: ; (This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB) ; Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT; Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT; Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT; Wash slides in 3 changes of xylene - 1 minute each @ RT; Apply coverslip
• Preparation and Storage: +4°C or -20°C, Avoid repeated freezing and thawing.

ELISA

(Anti-Insulin Antibody - ELISA. ELISA Results of Mab anti-Insulin antibody tested against human insulin by ELISA. Each well was coated with 0.1ug of conjugate. The starting concentration of antibody in the dilution series was 10 ug/ml. Each point on the Y-axis represents a 3-fold dilution. The midpoint of the titration curve represents approximately 5 ng/ml antibody or a 1:200000 dilution from the stock concentration. HRP conjugated Gt-a-Mouse IgG H&L and TMB substrate were used for detection.)

DB (Dot Blot)

(Anti-Insulin Antibody - Dot Blot. Dot blotting. Mab anti-Insulin antibody (clone 2D11. H5) is shown to detect human insulin by dot blot. Each dot blot represents 1 ul of non-denatured human insulin at various dilutions starting at 1.0 ug/ml spotted on to nitrocellulose. A 1:400 dilution of Mab anti-Insulin is used for 2 hour followed by detection using a 1:5000 dilution of IRDyeTM800 conjugated Goat-a-Mouse IgG [H&L] (reacted for 45 min at room temperature and visualization using the Odyssey Infrared Imaging System developed by LI-COR. Other detection systems will yield similar results. IRDye is a trademark of LI-COR, Inc.)

IHC (Immunohistochemistry)

(Anti-Insulin antibody IHC of human pancreas. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 5 ug/ml.)

Related Product Information for anti-INS antibody

Immunohistochemistry: MBS245024 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for MBS245024 was determined to be 5 ug/ml.

Product Categories/Family for anti-INS antibody

Family: Hormone

NCBI and Uniprot Product Information

• NCBI GI #: 4557671
• NCBI GeneID: 3630
• NCBI Accession #: NP_000198.1
• NCBI GenBank Nucleotide #: NM_000207.2
• UniProt Accession #: P01308; Q5EEX2
• NCBI Official Full Name: insulin preproprotein
• NCBI Official Synonym Full Names: insulin
• NCBI Official Symbol: INS
• NCBI Official Synonym Symbols: IDDM; ILPR; IRDN; IDDM1; IDDM2; MODY10
• NCBI Protein Information: insulin; proinsulin; preproinsulin
• UniProt Protein Name: Insulin
• UniProt Gene Name: INS
• UniProt Entry Name: INS_HUMAN