Anti-FABP5 antibody (ab84028)
Key features and details
- Rabbit polyclonal to FABP5
- Suitable for: ICC/IF, IP, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-FABP5 antibody
See all FABP5 primary antibodies -
Description
Rabbit polyclonal to FABP5 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Sheep, Cow, Pig
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Immunogen
Synthetic peptide corresponding to Human FABP5 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab94388) -
Positive control
- WB: HeLa, PC-3, HepG2 and A431 whole cell lystes; Skin tissue lysate. ICC/IF: HeLa cells.
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General notes
This product was previously labelled as Fatty Acid Binding Protein 5
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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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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KO cell pellets
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab84028 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 ICC/IF Use a concentration of 5 µg/ml. IP Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 16 kDa (predicted molecular weight: 15 kDa). Target
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Function
High specificity for fatty acids. Highest affinity for C18 chain length. Decreasing the chain length or introducing double bonds reduces the affinity. May be involved in keratinocyte differentiation. -
Tissue specificity
Keratinocytes; highly expressed in psoriatic skin. -
Sequence similarities
Belongs to the calycin superfamily. Fatty-acid binding protein (FABP) family. -
Domain
Forms a beta-barrel structure that accommodates the hydrophobic ligand in its interior. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 2171 Human
- Omim: 605168 Human
- SwissProt: Q01469 Human
- Unigene: 408061 Human
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Alternative names
- CFABP antibody
- Cutaneous fatty acid binding protein antibody
- DA11 antibody
see all
Images
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Western blot - Anti-FABP5 antibody (ab84028)All lanes : Anti-FABP5 antibody (ab84028) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FABP5 knockout HeLa cell lysate
Lane 3 : PC-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab84028 observed at 17 kDa. Red - loading control ab8245 observed at 36 kDa.
ab84028 Anti-FABP5 antibody was shown to specifically react with FABP5 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265905 (knockout cell lysate ab257431) was used. Wild-type and FABP5 knockout samples were subjected to SDS-PAGE. ab84028 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-FABP5 antibody (ab84028)ICC/IF image of ab84028 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab84028, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2, MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml. -
Western blot - Anti-FABP5 antibody (ab84028)All lanes : Anti-FABP5 antibody (ab84028) at 1 µg/ml
Lane 1 : Human skin tissue lysate - total protein (ab30166)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes -
Immunoprecipitation - Anti-FABP5 antibody (ab84028)FABP5 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab84028.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 16kDa; FABP5.
Protocols
References (4)
ab84028 has been referenced in 4 publications.
- Zaatiti H et al. Tumorigenic proteins upregulated in the MYCN-amplified IMR-32 human neuroblastoma cells promote proliferation and migration. Int J Oncol 52:787-803 (2018). PubMed: 29328367
- Li M et al. Comparative iTRAQ proteomics revealed proteins associated with horn development in yak. Proteome Sci 16:14 (2018). PubMed: 30061793
- Lu JB et al. Altered epidermal fatty acid-binding protein expression in hepatocellular carcinoma predicts unfavorable outcomes. Cancer Manag Res 10:6275-6284 (2018). PubMed: 30538573
- Suojalehto H et al. Level of Fatty Acid Binding Protein 5 (FABP5) Is Increased in Sputum of Allergic Asthmatics and Links to Airway Remodeling and Inflammation. PLoS One 10:e0127003 (2015). PubMed: 26020772
Images
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Western blot - Anti-FABP5 antibody (ab84028)All lanes : Anti-FABP5 antibody (ab84028) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FABP5 knockout HeLa cell lysate
Lane 3 : PC-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab84028 observed at 17 kDa. Red - loading control ab8245 observed at 36 kDa.
ab84028 Anti-FABP5 antibody was shown to specifically react with FABP5 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265905 (knockout cell lysate ab257431) was used. Wild-type and FABP5 knockout samples were subjected to SDS-PAGE. ab84028 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-FABP5 antibody (ab84028)ICC/IF image of ab84028 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab84028, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2, MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml.
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Western blot - Anti-FABP5 antibody (ab84028)All lanes : Anti-FABP5 antibody (ab84028) at 1 µg/ml
Lane 1 : Human skin tissue lysate - total protein (ab30166)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes
-
Immunoprecipitation - Anti-FABP5 antibody (ab84028)
FABP5 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab84028.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 16kDa; FABP5.
