HNF1 Antibody - BSA Free
Novus Biologicals | Catalog # NBP1-33596
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Theoretical MW
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for HNF1 Antibody - BSA Free
Immunoprecipitation: HNF1 Antibody [NBP1-33596]
Immunoprecipitation: HNF1 Antibody [NBP1-33596] - Immunoprecipitation of HNF1 alpha protein from HepG2 whole cell extracts using 5 ug of HNF1 alpha antibody [N1N3]. Western blot analysis was performed using HNF1 alpha antibody [N1N3]. EasyBlot anti-Rabbit IgG was used as a secondary reagent.Western Blot: HNF1 Antibody [NBP1-33596] -
Western Blot: HNF1 Antibody [NBP1-33596] - (A) Analysis of CBP/p300 DNA binding activity after TGF beta treatment. qPCR analysis of chromatin immunoprecipitated with anti-CBP/p300 antibody from hepatocytes untreated or treated with TGF beta (for 5 h) was performed. HNF1 alpha consensus regions embedded in the indicated target gene promoters were analyzed. A HNF1 alpha nonbound region of Neurogenin 1 promoter was utilized as negative control. Data are normalized to total chromatin input & background (control immunoprecipitation with IgG) & expressed as % input. Mean ± SEM of qPCR data obtained in triplicate from three independent experiments are reported. (B)In vivo coimmunoprecipitation of HNF1 alpha & CBP/p300 in hepatocytes untreated or treated with TGF beta (for 5 h). Cells were lysed, immunoprecipitated with anti-HNF1 alpha antibody, & then analyzed by Western blotting with the indicated antibodies. As control, the immunoprecipitation with normal goat IgG was performed. TCE, total cell extracts. Densitometric analysis of WB data from three independent experiments is shown. (C) Analysis of HNF1 alpha protein PTMs following TGF beta treatment. Nuclear extracts from control hepatocytes (upper panel) & HNF1 alpha Myc-overexpressing hepatocytes treated for 3 h with TGF beta (lower panel) or left untreated (middle panel). Samples were separated by two-dimensional gel electrophoresis followed by Western blotting with anti-Myc-Tag antibody. HNF1 alpha -specific spots are indicated by the arrow. The appearance of multiple spots in TGF beta -treated hepatocytes can be observed. **p <0.05, **p<0.01. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31543815), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: HNF1 Antibody [NBP1-33596] -
Western Blot: HNF1 Antibody [NBP1-33596] - Mouse tissue extract (50 ug) was separated by 7.5% SDS-PAGE, and the membrane was blotted with HNF1 antibody [N1N3] (NBP1-33596) diluted at 1:2000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.Immunocytochemistry/ Immunofluorescence: HNF1 Antibody [NBP1-33596] -
Immunocytochemistry/ Immunofluorescence: HNF1 Antibody [NBP1-33596] - HNF1 antibody [N1N3] detects HNF1 protein at nucleus by immunofluorescent analysis.Sample: HepG2 cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HNF1 stained by HNF1 antibody [N1N3] (NBP1-33596) diluted at 1:500.
Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] diluted at 1:1000.
Scale bar= 10um.
Western Blot: HNF1 Antibody [NBP1-33596] -
Western Blot: HNF1 Antibody [NBP1-33596] - Various whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HNF1 antibody [N1N3] (NBP1-33596) diluted at 1:2000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.Western Blot: HNF1 Antibody [NBP1-33596] -
Western Blot: HNF1 Antibody [NBP1-33596] - Various whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HNF1 alpha antibody [N1N3] diluted at 1:2000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.Applications for HNF1 Antibody - BSA Free
Immunocytochemistry/ Immunofluorescence
Immunohistochemistry
Immunohistochemistry-Paraffin
Immunoprecipitation
Western Blot
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Background: HNF1
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Additional HNF1 Products
Product Documents for HNF1 Antibody - BSA Free
Certificate of Analysis
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Product Specific Notices for HNF1 Antibody - BSA Free
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.
Citations for HNF1 Antibody - BSA Free
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Immunoprecipitation Protocol
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for HNF1 Antibody - BSA Free
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Q: I would like to know if the HNF1A antibody (NBP1-33596) cross reacts with HNF1B protein. The immunogen for HNF1A is a recombinant fragment corresponding to a region within amino acids 35 and 268 of HNF1 alpha. That particular fragment has a very high homology with HNF1B, in some areas 100%. We have already purchased the HNF1A antibody and conducted experiments using it for immunohistochemistry based on the assumption that it is a specific antibody (as it is stated on the datasheet). I now understand that the specificity of the antibody is uncertain. It is imperative for the interpretation of our data to be certain that the claimed specificity HNF1A antibody is a tested certainty. Since we have spent money (out of a tight budget) and effort on the project we would like to further test the HNF1A antibody cross reactivity with HNF1B. Could you please help us? I could think of two ways. First if you could run cross-reactivity tests, or secondly, if you could provide us with aliquots of the HNF1B antibody and lysates enriched for HNF1A and HNF1B so we could test the specificity in Western blots and immunohistochemistry. Our budget is very tight and we cannot pay for full price only to test the cross-reactivity of the products. It is imperative for our work to exclude the possibility of cross-reaction between HNF1A and HNF1B. In order to do so we need aliquots of the HNF1B antibody and lysates enriched for HNF1A and HNF1B so we could test the specificity in Western blots and immunohistochemistry. Therefore, we do not wish a direct refund or replacement which you kindly offered. Instead, we would like to ask for a generous discount on your three products that we need (HNF1B antibody and lysates enriched for HNF1A and HNF1B) taking into account the situation and our limited budget. In order for the discount to be bigger we would like the quantity from each product to be less than usual since we just want to check the cross-reactivity issue. I would like to have a credit for the cost of the NBP1-33596 antibody, and use it to reduce the cost of the products I would like. You also said that you may be able to get me a discount on top of this.
A: Although we have not tested the cross-reactivity of antibody NBP1-33596 with HNF1B, I have performed a sequence comparison analysis for you, and HNF1A only has 54% homology with HNF1B (56% for isoform B), suggesting that the antibody is unlikely to bind to HNF1B.The website data sheet does state which portion of HNF1 alpha the antibody was raised against, so this information is available for our customers before they begin their experiments. HNF1 alpha was the protein this antibody was generated for, but cross-reactivity against other proteins was not tested. We do not state that the antibody will definitely not bind to HNF1 beta. However, I understand that the datasheet can be misinterpreted. Unfortunately, we are unable to carry out your requests, but we can offer you a refund or replacement. Please let me know how you wish to proceed. I am making enquiries into whether any of our antibodies are known to be specific to either HNF1B or HNF1A, and will let you know as soon as I have an answer. We do have overexpression lysates for HNF1A and HNF1B: NBL-11630 and NBL-11631. Unfortunately, we cannot change the pack size of any of our products. However, we will give you a credit for the cost of the NBP1-33596 antibody, and you can use this to reduce the cost of the products you would like. I may be able to get you a discount on top of this. Unfortunately, none of the HNF1A antibodies have been tested for cross-reactivity to HNF1B, and none of the HNF1B antibodies have been tested for cross-reactivity to HNF1A. Therefore, we do not have any antibodies that I can guarantee are specific to either HNF1A or HNF1B; please accept my apologies. Please could you let me know the PO of your original order with us? Or did you order via a distributor? Also, as I said in my previous email, we do not have an HNF1B antibody that has been tested for cross-reactivity against HNF1A. However, if you would still like one from us, I would recommend NBP1-89680, because the lab have let me know that the antigen of this antibody has 47% sequence identity to the HNF1A gene. This antibody shows nuclear staining of moderate to strong intensity in the gastrointestinal tract, renal tubules, pancreatic ducts and decidual cells; it shows no positivity in hepatocytes. 'Although HNF-1 alpha, but not HNF-1 beta, is expressed in the liver, HNF-1 alpha and HNF-1 beta are coexpressed in the murine Hepa1A cell line and in the mammalian kidney where a subset of hepatocyte genes are expressed', accor