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.

Name: HNF1 Antibody
Brand: Novus Biologicals
SKU: NBP1-33596

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

HNF1 Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-33596

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Product Datasheet / COA

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Predicted:

Bovine (98%), Chicken (95%), Porcine (99%). Backed by our 100% Guarantee.

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, SDS-Page, Western Blot

Cited:

Immunocytochemistry/ Immunofluorescence, SDS-Page, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

View all HNF1 Antibodies »

Product Specifications

Immunogen

Recombinant protein encompassing a sequence within the center region of human HNF1. The exact sequence is proprietary.

Reactivity Notes

Xenopus laevis (85%).

Localization

Nucleus

Specificity

human HNF-1 alpha protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

67 kDa.
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

Western Blot: HNF1 Antibody [NBP1-33596]

Western Blot: HNF1 Antibody [NBP1-33596] - Analysis of HNF1a transcriptional activity in hepatocytes following transforming growth factor B (TGFB) treatment. (B) Western blot analysis for HNF1a in protein extracts from hepatocytes in a time-course experiment following TGFB treatment. a-Glyceraldehyde 3-phosphate dehydrogenase was used as loading control. Magnification 20x. Image collected and cropped by CiteAb from the following publication (frontiersin.org/article/10.3389/fphar.2019.00942/full) licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: HNF1 Antibody [NBP1-33596]

Immunocytochemistry/Immunofluorescence: HNF1 Antibody [NBP1-33596] - TGFB overrides HNF1a constitutive expression. Immunofluorescence analysis of hepatocytes transfected with pLCPX-HNF1aMyc (HNF1) or the empty vector (CTR), treated with TGFB for 24 hr or left untreated (NT). . Cells were stained with anti-E-cadherin (green) or anti-HNF1a antibody (red). Magnification 20x. Image collected and cropped by CiteAb from the following publication (frontiersin.org/article/10.3389/fphar.2019.00942/full) licensed under a CC-BY license.

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 1:5000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596]

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] - Paraffin-embedded A549 Xenograft, using antibody at 1:100 dilution.

Western Blot: HNF1 Antibody [NBP1-33596]

Western Blot: HNF1 Antibody [NBP1-33596] - Rat tissue extracts (50 ug) was separated by 7.5% SDS-PAGE, and the membrane was blotted with HNF1 alpha antibody [N1N3] diluted at 1:1000.

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596]

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] - detects HNF1 alpha protein at nucleus by immunohistochemical analysis.Sample: Paraffin-embedded mouse colon.HNF1 alpha stained by HNF1 alpha antibody [N1N3] diluted at 1:500.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

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 alphaMyc-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.

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] -

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] - HNF1 antibody [N1N3] detects HNF1 protein at cytoplasm and nucleus by immunohistochemical analysis.
Sample: Paraffin-embedded mouse colon.
HNF1 stained by HNF1 antibody [N1N3] (NBP1-33596) diluted at 1:500.
Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] -

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] - HNF1 antibody [N1N3] detects HNF1 protein at nucleus by immunohistochemical analysis.
Sample: Paraffin-embedded mouse intestine.
HNF1 stained by HNF1 antibody [N1N3] (NBP1-33596) diluted at 1:500.
Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

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.

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Applications for HNF1 Antibody

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Immunohistochemistry

1:100-1:1000

Immunohistochemistry-Paraffin

1:100-1:1000

Immunoprecipitation

1:100-1:500

Western Blot

1:1000-1:10000

Application Notes

Use in SDS-Page reported in scientific literature (PMID:31543815).

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

PBS (pH 7), 20% Glycerol

Preservative

0.025% Proclin 300

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: HNF1

HNF1 is a transcription factor required for the expression of several liver-specific genes. The protein functions as a homodimer and binds to the inverted palindrome 5'-GTTAATNATTAAC-3'. Defects in this gene are a cause of maturity onset diabetes of the young type 3 (MODY3) and also can result in the appearance of hepatic adenomas. Alternative splicing results in multiple transcript variants encoding different isoforms.

Alternate Names

HNF1 homeobox A, IDDM20, LFB1transcription factor 1, hepatic; LF-B1, hepatic nuclear factor (HNF1), albuminproximal factor, MODY3, TCF1hepatic, Transcription factor 1

Entrez Gene IDs

6927 (Human)

Gene Symbol

HNF1A

UniProt

P20823 (Human)

Additional HNF1 Products

Product Documents for HNF1 Antibody

Product Datasheets

Download Product Datasheets

COA

Product Specific Notices for HNF1 Antibody

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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HNF1 Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-33596

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Predicted:

Applications

Validated:

Cited:

Label

Antibody Source

Concentration

View all HNF1 Antibodies »

Product Specifications

Immunogen

Reactivity Notes

Localization

Specificity

Clonality

Host

Isotype

Theoretical MW

Scientific Data Images for HNF1 Antibody

Western Blot: HNF1 Antibody [NBP1-33596]

Immunocytochemistry/ Immunofluorescence: HNF1 Antibody [NBP1-33596]

Western Blot: HNF1 Antibody [NBP1-33596]

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596]

Western Blot: HNF1 Antibody [NBP1-33596]

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596]

Immunoprecipitation: HNF1 Antibody [NBP1-33596]

Western Blot: HNF1 Antibody [NBP1-33596] -

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] -

Immunohistochemistry-Paraffin: HNF1 Antibody [NBP1-33596] -

Western Blot: HNF1 Antibody [NBP1-33596] -

Immunocytochemistry/ Immunofluorescence: HNF1 Antibody [NBP1-33596] -

Western Blot: HNF1 Antibody [NBP1-33596] -

Western Blot: HNF1 Antibody [NBP1-33596] -

Applications for HNF1 Antibody

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Paraffin

Immunoprecipitation

Western Blot

Formulation, Preparation, and Storage

Purification

Formulation

Preservative

Concentration

Shipping

Stability & Storage

Background: HNF1

Alternate Names

Entrez Gene IDs

Gene Symbol

UniProt

Additional HNF1 Products

Product Documents for HNF1 Antibody

Product Datasheets

COA

Certificate of Analysis

Product Specific Notices for HNF1 Antibody