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Proinsulin Antibody (3A1)

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Product Details

Summary
Reactivity Hu, Bv(-), Po(-)Species Glossary
Applications IHC, IHC-Fr, S-ELISA
Clone
3A1
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

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Proinsulin Antibody (3A1) Summary

Immunogen
Hybridoma clone has been derived from hybridization of X63-Ag8-653 myeloma cells with spleen cells of Balb/c mice immunized with purified human proinsulin.
Specificity
Kd for this is >1.1 x 10^-8M. Reacts with the site of connection of C-peptide and C-end of B-chain of proinsulin molecule. Does not cross-react with human, bovine and porcine insulin, bovine and porcine proinsulin, and human C-peptide.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
INS
Purity
Protein A purified
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Applications/Dilutions

Dilutions
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Sandwich ELISA 1:100-1:2000
Application Notes
Recommended pairs for sandwich immunoassay (capture-detection): NB110-2558 - NB600-1448

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS (pH 7.4)
Preservative
0.09% Sodium Azide
Purity
Protein A purified

Alternate Names for Proinsulin Antibody (3A1)

  • IDDM2
  • ILPR
  • insulin
  • IRDN
  • MODY10
  • Proinsulin

Background

A precursor of insulin, with a molecular weight of 8,000 to 10,000. It has minimum hormonal activity and is converted to insulin by removal of the connecting C peptide, leaving the two (A and B) chains. From every molecule of proinsulin, one molecule of insulin plus one molecule of C Peptide are produced.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Glucagon was detected in immersion fixed beta TC-6 mouse beta cell insulinoma cell line using 10 µg/mL Human/Mouse Glucagon Monoclonal Antibody (Catalog # MAB1249) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
beta TC‑6 mouse beta cell insulinoma cell line was stained with Mouse Anti-Human/Mouse Glucagon Monoclonal Antibody (Catalog # MAB1249, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
(3 Reviews)
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IGF-I/IGF-1
[Unconjugated]
291-G1
1 μg/lane of Recombinant Human IGF-I/IGF-1 was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 7 kDa.
Recombinant Human IGF-I/IGF-1 (Catalog # 291-G1) stimulates proliferation in the MCF-7 human breast cancer cell line. The ED50 for this effect is 0.3‑1.5 ng/mL.
MALDI-TOF analysis of Recombinant Human IGF-I/IGF-1.The major peak corresponds to the calculated molecular mass, 7650 Da. The minor peak at 7873 is a matrix-associated artifact of the MALDI-TOF.
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Western blot shows lysates of human, mouse, and rat pituitary gland tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Growth Hormone Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1067) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Growth Hormone at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Recombinant Human Growth Hormone protein was serially diluted 2-fold and captured by Mouse Anti-Human Growth Hormone Monoclonal Antibody (Catalog # MAB10671) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human/Mouse/Rat Growth Hormone Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1067) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Recombinant Human Growth Hormone (Catalog # 1067-GH) stimulates proliferation in the Nb2‑11 rat lymphoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Growth Hormone (0.2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Mouse/Rat Growth Hormone Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1067). The ND50 is typically 1.5-7.5 ng/mL.
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(8 Publications)
Species: Hu, Mu, Rt
Applications: ELISA, IHC, Neut, WB
Insulin Receptor
beta Antibody
NBP2-12793
Immunohistochemistry-Paraffin: Insulin Receptor beta Antibody [NBP2-12793] - Staining shows expression of insulin receptor beta in the Cortex in mouse brain. Dilution 1:500. Image submitted by a verified customer review.
Immunoprecipitation: Insulin Receptor beta Antibody [NBP2-12793] - Samples: Whole cell lysate from HeLa (15 and 50 ug for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 ug), Jurkat (J; 50 ug) and mouse NIH3T3 (M; 50 ug) cells. Antibodies: Affinity purified rabbit anti-Insulin Receptor Beta antibody NBP2-12793 used for WB at 0.1 ug/ml (A) and 1 ug/ml (B) and used for IP at 6 ug/mg lysate. Insulin Receptor Beta was also immunoprecipitated by rabbit anti-Insulin Receptor Beta antibody, which recognizes an upstream epitope. Detection: Chemiluminescence with exposure times of 30 seconds (A and B).
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Adiponectin/Acrp30
[HRP]
DRP300
(6 Reviews)
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TNF-alpha
[Unconjugated]
210-TA
1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
Recombinant Human TNF-alpha (Catalog # 210‑TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25‑100 pg/mL.
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(702 Publications)
Species: Hu
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Growth Hormone
Antibody [Unconjugate
AF1067
Western blot shows lysates of human, mouse, and rat pituitary gland tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Growth Hormone Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1067) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Growth Hormone at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Somatostatin
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NBP2-37447
Immunofluorescence: Somatostatin Antibody (7G5) [NBP2-37447] - Paraffin-embedded human adult pancreas tissue. pH 9 antigen retrieval. Image from verified customer review.
Immunohistochemistry-Paraffin: Somatostatin Antibody (7G5) [NBP2-37447] - Analysis of lung cancer tissues using SST mouse mAb with DAB staining.
ELISA: Somatostatin Antibody (7G5) [NBP2-37447] - Red: Control Antigen (100ng); Purple: Antigen (10ng); Green: Antigen (50ng); Blue: Antigen (100ng);
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Glut4 Antibody -
BSA Free
NBP1-49533
Flow (Intracellular): Glut4 Antibody [NBP1-49533] - An intracellular stain was performed on HepG2 with NBP1-49533 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.
Western Blot: Glut4 Antibody [NBP1-49533] - Total protein from 3T3-L1 mouse embryonic fibroblast adipose-like cell line, separated by 4-12% SDS-PAGE, transferred to nitrocellulose membrane and blocked in 5% non-fat milk for 1h at room temperature. The membrane was probed with anti-Glut4 0.5 ug/ml in non-fat milk. Undiffer: undifferentiated; Differ: Differentiated. Image from verified customer review.
Western Blot: Glut4 Antibody [NBP1-49533] - Glycolysis-related genes and proteins were upregulated in differentiated podocytes. Representative blot images of glycolysis-related proteins (n =3). d All proteins were normalized to tubulin and compared to UDPs. *P < 0.05, **P < 0.01, determined by t test. Data are shown as the means +/- SD. Image collected and cropped by CiteAb from the following publication (http://www.nature.com/articles/s41419-020-2481-5), licensed under a CC-BY licence.
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hydrolase Antibody
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Western Blot: gamma-glutamyl hydrolase Antibody [NBP1-90927] - Analysis in control (vector only transfected HEK293T lysate) and GGH over-expression lysate (Co-expressed with a C-terminal myc-DDK tag (3.1 kDa) in mammalian HEK293T cells).
Immunohistochemistry-Paraffin: gamma-glutamyl hydrolase Antibody [NBP1-90927] - Staining of human kidney shows strong granular cytoplasmic positivity in cells in tubules.
Immunohistochemistry-Paraffin: gamma-glutamyl hydrolase Antibody [NBP1-90927] - Staining of human testis shows strong granular cytoplasmic positivity in Leydig cells.
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C-Reactive
Protein/CRP [HRP]
DCRP00
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EGF [Unconjugated]
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1 µg/lane of Recombinant Human EGF was resolved with SDS-PAGE  and visualized by silver staining under reducing (R) conditions, showing a single band at 6 kDa.
Recombinant Human EGF (Catalog # 236‑EG) stimulates cell proliferation of the Balb/3T3 mouse embryonic fibroblast cell line. The ED50 for this effect is 20‑100 pg/mL.
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PPAR gamma/NR1C3
Antibody - BSA Free
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Western Blot: PPAR gamma/NR1C3 Antibody [NBP2-22106] - Total protein from 3T3-L1 mouse embryonic fibroblast adipose-like cell line, separated by 4-12% SDS-PAGE, transferred to nitrocellulose membrane and blocked in 5% non-fat milk for 1h at room temperature. The membrane was probed with anti-PPAR gamma 1:800 in non-fat milk. Lane 2-7 increasing protein concentration, undifferentiated adipocytes. Lane 8-13 increasing protein concentration, differentiated adipocytes. This image was submitted via customer review.
Immunohistochemistry-Paraffin: PPAR gamma Antibody [NBP2-22106] - IHC analysis of PPAR gamma in mouse liver.
Western Blot: PPAR gamma Antibody [NBP2-22106] - Western blot analysis of PPAR gamma in 1. human adipose and 2. human adrenal lysate.
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IL-6 [HRP]
D6050
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AKT [p Ser473]
Antibody - Pan Specif
AF887
Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (Catalog # 120‑HD) for 15 minutes and MCF‑7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (Catalog # 291‑G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line and A549 human lung carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF‑I (Catalog # 291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Serum-starved MCF‑7 human breast cancer cells were unstimulated (light orange filled histogram) or activated with 100 ng/mL Recombinant human IGF-1 (Catalog # 291-G1, dark orange filled histogram) for 15 minutes, then stained with Rabbit Anti-Human/Mouse/Rat Phospho‑Akt (S473) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF887) or control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
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POMC Antibody
NB100-1533
Western Blot: POMC Antibody [NB100-1533] - Staining of NIH3T3 cell lysate with antibody at 0.3 ug/mL (A) and Rat Brain cell lysate with antibody at 2 ug/mL (B). 35 ug protein in RIPA buffer. Detected by chemiluminescence.
Flow Cytometry: POMC Antibody [NB100-1533] - Flow cytometric analysis of paraformaldehyde fixed A431 cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (1 ug/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.
Immunohistochemistry: POMC Antibody [NB100-1533] - Representative immunofluorescence images of (B) POMC as a prohormone (n = 5-6 each) in the ARC of mice approximately 5 to 10 weeks of age. Image collected and cropped by CiteAb from the following publication (jci.org/articles/view/96420), licensed under a CC-BY licence.
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Species: Hu, Mu, Po, Rt, Sh
Applications: Flow, ICC/IF, IHC, IHC-P, PEP-ELISA, WB
Albumin Antibody
(188835) [Unconjugate ...
MAB1455
Simple Western lane view shows lysates of human serum, loaded at 1:25000. A specific band was detected for Albumin at approximately 64 kDa (as indicated) using 1 µg/mL of Mouse Anti-Human Serum Albumin Monoclonal Antibody (Catalog # MAB1455) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Albumin was detected in immersion fixed BG01V human embryonic stem cells differentiated to hepatocytes using Mouse Anti-Human Serum Albumin Monoclonal Antibody (Catalog # MAB1455) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Cells were co-stained using Sheep Anti-Human CEBP alpha (Catalog # AF7094) and NorthernLights™ 493-conjugated Anti-Sheep IgG Secondary Antibody (green, Catalog # NL012). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Albumin was detected in immersion fixed paraffin-embedded sections of human liver using Mouse Anti-Human Serum Albumin Monoclonal Antibody (Catalog # MAB1455) at 0.1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
(5 Reviews)
(28 Publications)
Species: Hu
Applications: CyTOF-ready, Dual ISH-IHC, ICC, IHC, ICFlow, Simple Western, WB
Prolactin Antibody
[Unconjugated]
AF682
Recombinant Human Prolactin (Catalog # 682-PL) stimulates proliferation in the Nb2‑11 rat lymphoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Prolactin (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682). The ND50 is typically 0.02-0.05 µg/mL.
Prolactin was detected in immersion fixed paraffin-embedded sections of human testis using Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of sperm cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Western blot shows lysates of human pituitary tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Prolactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF682) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Prolactin at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
(6 Publications)
Species: Hu
Applications: ELISA(Cap), ELISA(Det), ELISA(Sta), IHC, Neut, WB
Thyroglobulin
Antibody (2H11 + 6E1)
NBP2-34294
Immunohistochemistry-Paraffin: Thyroglobulin Antibody (2H11 + 6E1) [NBP2-34294] - Analysis using Azide/BSA FREE version of NBP2-34294. Human Thyroid stained with Thyroglobulin Ab (2H11 + 6E1).
Immunohistochemistry: Thyroglobulin Antibody (2H11 + 6E1) [NBP2-34294] - Imaging of Mouse stomach tissue. Positive staining can only be observed inside the thyroid cells and the follicle in thyroid tissue. This image was submitted via customer Review.
Immunohistochemistry-Paraffin: Thyroglobulin Antibody (2H11 + 6E1) [NBP2-34294] - Thyroglobulin staining in thyroid tissue of normal C57 mouse (Red). Dilution is 1:100. IHC-P image submitted by a verified customer review.
(3 Reviews)
(1 Publication)
Species: Hu, Mu, Rt
Applications: Flow, IHC, IHC-P

Publications for Proinsulin Antibody (NB110-2558) (0)

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Product General Protocols

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FAQs for Proinsulin Antibody (NB110-2558). (Showing 1 - 1 of 1 FAQ).

  1. Looking for good proinsulin antibody for human fresh frozen samples. Insulin causes smearing.
    • Thank you for your interest in our products. We only have one proinsulin antibody that is validated in IHC frozen samples. The validation was done with fixed frozen sections, not fresh frozen; however, we will guarantee this antibody for you. This is a licensed product and the validation of this product has been done by the inventor of the clone. Unfortunately, we do not have any images on the website.

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Bioinformatics

Gene Symbol INS
Entrez