Recombinant Anti-TGF beta 1 antibody [EPR21143] (ab215715)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21143] to TGF beta 1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TGF beta 1 antibody [EPR21143]
See all TGF beta 1 primary antibodies -
Description
Rabbit monoclonal [EPR21143] to TGF beta 1 -
Host species
Rabbit -
Specificity
For testing samples with low expression level of TGF beta 1, we recommend ab179695 which could give stronger signal. Loading larger amount of lysate or lower antibody dilution would also help.
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Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NIH/3T3, L-929, HeLa, A549, HL-60, RAW 264.7, Wild-type A549, K562 and SH-SY5Y whole cell lysates; Mouse and Rat spleen lysate; Mouse heart tissue lysate; C6 cell lysate. IHC-P: Human thrombocytosis tissue; Mouse and rat spleen tissues.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21143 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215715 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (1) |
1/1000. Predicted molecular weight: 44 kDa.
Actual band: 13 kDa and 44 kDa |
IHC-P | (4) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Predicted molecular weight: 44 kDa. Actual band: 13 kDa and 44 kDa |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. Many cells synthesize TGFB1 and have specific receptors for it. It positively and negatively regulates many other growth factors. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts. -
Tissue specificity
Highly expressed in bone. Abundantly expressed in articular cartilage and chondrocytes and is increased in osteoarthritis (OA). Co-localizes with ASPN in chondrocytes within OA lesions of articular cartilage. -
Involvement in disease
Defects in TGFB1 are the cause of Camurati-Engelmann disease (CE) [MIM:131300]; also known as progressive diaphyseal dysplasia 1 (DPD1). CE is an autosomal dominant disorder characterized by hyperostosis and sclerosis of the diaphyses of long bones. The disease typically presents in early childhood with pain, muscular weakness and waddling gait, and in some cases other features such as exophthalmos, facial paralysis, hearing difficulties and loss of vision. -
Sequence similarities
Belongs to the TGF-beta family. -
Post-translational
modificationsGlycosylated.
The precursor is cleaved into mature TGF-beta-1 and LAP, which remains non-covalently linked to mature TGF-beta-1 rendering it inactive. -
Cellular localization
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 7040 Human
- Entrez Gene: 21803 Mouse
- Entrez Gene: 59086 Rat
- Omim: 190180 Human
- SwissProt: P01137 Human
- SwissProt: P04202 Mouse
- SwissProt: P17246 Rat
- Unigene: 645227 Human
see all -
Alternative names
- Cartilage-inducing factor antibody
- CED antibody
- Differentiation inhibiting factor antibody
see all
Images
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All lanes : Anti-TGF beta 1 antibody [EPR21143] (ab215715) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2 : HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg
Lane 3 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 6 : Mouse spleen tissue lysate
Lane 7 : Rat spleen tissue lysate
Lane 8 : Mouse heart tissue lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 12,44 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
For testing samples with low expression level of TGF beta 1, we recommend ab179695 which could give stronger signal. Loading larger amount of lysate or lower antibody dilution would also help.
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All lanes : Anti-TGF beta 1 antibody [EPR21143] (ab215715) at 1/1000 dilution
Lane 1 : Wild-type HeLa whole cell lysate
Lane 2 : TGFB1 knockout HeLa whole cell lysate
Lane 3 : A549 whole cell lysate
Lane 4 : Mouse spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 44 kDa
Observed band size: 13,44 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab215715 observed at 13 and 44 kDa. Red - loading control, ab7291, observed at 50 kDa.
ab215715 was shown to specifically react with in wild-type HeLa cells as signal was lost in TGFB1 knockout cells. Wild-type and TGFB1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab215715 and ab7291 (Mouse anti-tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-TGF beta 1 antibody [EPR21143] (ab215715) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : TGFB1 knockout A549 cell lysate
Lane 3 : K562 cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab215715 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab215715 was shown to react with TGF beta 1 in wild-type A549 cells in Western blot with loss of signal observed in TGFB1 knockout cell line ab269509 (TGFB1 knockout cell lysate ab269671). Wild-type A549 and TGFB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab215715 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGF beta 1 antibody [EPR21143] (ab215715)
Immunohistochemical analysis of paraffin-embedded human thrombocytosis tissue labeling TGF beta 1 with ab215715 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining of megakaryocytes in human thrombocytosis (PMID: 25305163). Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGF beta 1 antibody [EPR21143] (ab215715)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling TGF beta 1 with ab215715 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of megakaryocytes and platelets in rat spleen (PMID: 25305163). Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGF beta 1 antibody [EPR21143] (ab215715)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling TGF beta 1 with ab215715 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of megakaryocytes and platelets in mouse spleen (PMID: 25305163). Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
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All lanes : Anti-TGF beta 1 antibody [EPR21143] (ab215715) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
Lane 2 : L-929 (mouse connective tissue fibroblast cell line) whole cell lysate at 20 µg
Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4 : HL-60 (human promyelocytic leukemia cell line) whole cell lysate at 20 µg
Lane 5 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
Lane 6 : Mouse spleen lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 12,44 kDa why is the actual band size different from the predicted?Exposure times: Lanes 1-4: 3 minutes; Lane 5: 41 seconds; Lane 6: 24 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular mass observed is consistent with the literature (PMID 2139036).
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (134)
ab215715 has been referenced in 134 publications.
- Zheng D et al. Biomimetic nanoparticles drive the mechanism understanding of shear-wave elasticity stiffness in triple negative breast cancers to predict clinical treatment. Bioact Mater 22:567-587 (2023). PubMed: 36382024
- Wu J et al. CD147 contributes to SARS-CoV-2-induced pulmonary fibrosis. Signal Transduct Target Ther 7:382 (2022). PubMed: 36424379
- Lu Y et al. Tocotrienol-Rich Fractions Offer Potential to Suppress Pulmonary Fibrosis Progression. Int J Mol Sci 23:N/A (2022). PubMed: 36430808
- Zhou W et al. Sophocarpine Alleviates Isoproterenol-Induced Kidney Injury by Suppressing Inflammation, Apoptosis, Oxidative Stress and Fibrosis. Molecules 27:N/A (2022). PubMed: 36431969
- Gong P et al. Mesenchymal stem cells alleviate systemic sclerosis by inhibiting the recruitment of pathogenic macrophages. Cell Death Discov 8:466 (2022). PubMed: 36435837