Anti-SREBP1 antibody (ab28481)

Rabbit polyclonal SREBP1 antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 24 publication(s). Independently reviewed in 9 review(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to SREBP1
  • Host species
    Rabbit
  • Specificity
    ab28481 recognises SREBP1.
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    MLQLINNQDSDFPGLF

    , corresponding to amino acids 32-47 of Mouse SREBP 1

  • Positive control
    • Mouse and rat liver. Rat kidney.

Applications

Our Abpromise guarantee covers the use of ab28481 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
WB 1/500 - 1/5000. Can be blocked with Mouse SREBP1 peptide (ab31099).

Detects an ~68 and 120 kDa protein representing SREBP1 in mouse and rat liver samples as well as rat kidney samples. A predominant band at ~68 kDa (active cleaved site) is seen and a band at ~120 kDa (inactive precursor) may not be seen or it may be diminished.

ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.
IHC-P 1/2000.

Target

  • Function
    Transcriptional activator required for lipid homeostasis. Regulates transcription of the LDL receptor gene as well as the fatty acid and to a lesser degree the cholesterol synthesis pathway (By similarity). Binds to the sterol regulatory element 1 (SRE-1) (5'-ATCACCCCAC-3'). Has dual sequence specificity binding to both an E-box motif (5'-ATCACGTGA-3') and to SRE-1 (5'-ATCACCCCAC-3').
  • Tissue specificity
    Expressed in a wide variety of tissues, most abundant in liver and adrenal gland. In fetal tissues lung and liver shows highest expression. Isoform SREBP-1C predominates in liver, adrenal gland and ovary, whereas isoform SREBP-1A predominates in hepatoma cell lines. Isoform SREBP-1A and isoform SREBP-1C are found in kidney, brain, white fat, and muscle.
  • Sequence similarities
    Belongs to the SREBP family.
    Contains 1 basic helix-loop-helix (bHLH) domain.
  • Post-translational
    modifications
    At low cholesterol the SCAP/SREBP complex is recruited into COPII vesicles for export from the ER. In the Golgi complex SREBPs are cleaved sequentially by site-1 and site-2 protease. The first cleavage by site-1 protease occurs within the luminal loop, the second cleavage by site-2 protease occurs within the first transmembrane domain and releases the transcription factor from the Golgi membrane. Apoptosis triggers cleavage by the cysteine proteases caspase-3 and caspase-7.
    Phosphorylated by AMPK, leading to suppress protein processing and nuclear translocation, and repress target gene expression. Phosphorylation at Ser-402 by SIK1 represses activity possibly by inhibiting DNA-binding.
  • Cellular localization
    Nucleus and Endoplasmic reticulum membrane. Golgi apparatus membrane. Cytoplasmic vesicle > COPII-coated vesicle membrane. Moves from the endoplasmic reticulum to the Golgi in the absence of sterols.
  • Information by UniProt
  • Database links
  • Alternative names
    • ADD 1 antibody
    • bHLHd1 antibody
    • Class D basic helix-loop-helix protein 1 antibody
    • D630008H06 antibody
    • Processed sterol regulatory element-binding protein 1 antibody
    • SRBP1_HUMAN antibody
    • SREBF 1 antibody
    • SREBF1 antibody
    • SREBP 1 antibody
    • SREBP 1c antibody
    • SREBP-1 antibody
    • SREBP1 antibody
    • Sterol regulatory element binding protein 1 antibody
    • Sterol Regulatory Element Binding Transcription Factor 1 / Protein 1 antibody
    • Sterol regulatory element binding transcription factor 1 antibody
    • Sterol regulatory element-binding transcription factor 1 antibody
    see all

Images

  • All lanes : Anti-SREBP1 antibody (ab28481) at 1/1000 dilution

    Lane 1 : Mouse Liver Whole Cell Lysate
    Lane 2 : Rat Liver Whole Cell Lysate
    Lane 3 : HepG2 whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP-Conjugate
  • Immunocytochemical analysis of formalin-fixed C2C12 cell lines using immunofluorescence to label SREBP1 with ab28481 at a concentration of 1/100 in 3% BSA-PBS and incubated overnight in a humid environment at 4°C. Prior to labelling, cells were permeablised with 0.1% Triton X-100 in TBS for between 5 and 10 minutes, they were subsequently blocked with 3% BSA-PBS for 30 minutes at room temperature. The secondary used was a DyLight® conjugate (green) and was incubated at room temperature in the dark. The cells were counterstained with DAPI against DNA labelling the nucear compartments blue and a red fluorescent phalloidin stain against F-Actin. Magnification was 60X
    The left image is a negative control in the absence of ab28481, the right image is in the prescence of ab28481, the secondary and counterstains.

  • ab28481 staining SREBP1 in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 15 minutes at 25°C. Samples were incubated with primary antibody (1/2000 in goat serum) for 12 hours at 4°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunocytochemical analysis of formalin-fixed HepG2 cell lines using immunofluorescence to label SREBP1 with ab28481 at a concentration of 1/100 in 3% BSA-PBS and incubated overnight in a humid environment at 4°C. Prior to labelling, cells were permeablised with 0.1% Triton X-100 in TBS for between 5 and 10 minutes, they were subsequently blocked with 3% BSA-PBS for 30 minutes at room temperature. The secondary used was a DyLight® conjugate (green) and was incubated at room temperature in the dark. The cells were counterstained with DAPI against DNA labelling the nucear compartments blue and a red fluorescent phalloidin stain against F-Actin. Magnification was 60X
    The left image is a negative control in the absence of ab28481, the right image is in the prescence of ab28481, the secondary and counterstains.

  • Immunocytochemical analysis of formalin-fixed NIH 3T3 cell lines using immunofluorescence to label SREBP1 with ab28481 at a concentration of 1/100 in 3% BSA-PBS and incubated overnight in a humid environment at 4°C. Prior to labelling, cells were permeablised with 0.1% Triton X-100 in TBS for between 5 and 10 minutes, they were subsequently blocked with 3% BSA-PBS for 30 minutes at room temperature. The secondary used was a DyLight® conjugate (green) and was incubated at room temperature in the dark. The cells were counterstained with DAPI against DNA labelling the nucear compartments blue and a red fluorescent phalloidin stain against F-Actin. Magnification is 60X
    The left image is a negative control in the absence of ab28481, the right image is in the prescence of ab28481, the secondary and counterstains.

  • ICC/IF image of ab28481 stained HepG2 cells. The cells were 10% formaldehyde 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 (ab28481, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, goat anti rabbit DyLight® 488 IgG (H+L) used at a 1/250 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.

  • Immunohistochemistry (Frozen sections) analysis of mouse liver tissue labeling SREBP1 with ab28481 at 1/200 dilution. Tissue was fixed in acetone followed by blocking with 10% serum for 1 hour at 21°C. The tissue was stained with ab28481 at 1/200 for 12 hours in PBS+0,1% Triton X at 4°C. A polyclonal goat anti-rabbit Alexa Fluor® 594 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Immunohistochemistry (Frozen sections) analysis of rat liver tissue labeling SREBP1 with ab28481 at 1/200 dilution. Tissue was fixed in acetone followed by blocking with 10% serum for 1 hour at 21°C. The tissue was stained with ab28481 at 1/200 for 12 hours in PBS+0,1% Triton X at 4°C. A polyclonal goat anti-rabbit Alexa Fluor® 594 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Immunohistochemistry (Frozen sections) analysis of rat liver tissue labeling SREBP1 with ab28481 at 1/200 dilution. Tissue was fixed in acetone followed by blocking with 10% serum for 1 hour at 21°C. The tissue was stained with ab28481 at 1/200 for 14 hours in PBS+0,1% Triton X at 4°C. A polyclonal goat anti-rabbit Alexa Fluor® 594 secondary antibody was used at 1/1000 dilution.

    See Abreview

References

This product has been referenced in:
  • Xu MX  et al. Prolonged PM2.5 exposure elevates risk of oxidative stress-driven nonalcoholic fatty liver disease by triggering increase of dyslipidemia. Free Radic Biol Med 130:542-556 (2019). Read more (PubMed: 30465824) »
  • Dumolt JH  et al. Malprogramming of Hepatic Lipid Metabolism due to Excessive Early Cholesterol Exposure in Adult Progeny. Mol Nutr Food Res 63:e1800563 (2019). Read more (PubMed: 30447138) »
See all 28 Publications for this product

Customer reviews and Q&As

11-14 of 14 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Liver)
Loading amount
30 µg
Specification
Liver
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3

Abcam user community

Verified customer

Submitted Jun 18 2007

Answer

Thank you for your enquiry. We have not tested this product to determine whether it recognizes the C isoform specifically. Based on sequence similarity I would expect that it would recognize SREBP-1c (conserved sequence). Unfortunately we are not aware of publications where this antibody has been used. Please let me know if I can be of further assistance.

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Answer

Thank you for your enquiry. To our knowledge this antibody has not been applied in a publication. With regards the recognition of SREBP-1c by this antibody, I am in touch with the lab to determine whether this has been tested and will get back to you shortly. I appreciate your patience in this matter.

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Answer

Thank you for your enquiry. This antibody detects an ~68 and 120 kDa protein representing SREBP 1 in mouse and rat liver samples as well as rat kidney samples. A predominant band at ~68 kDa (active cleaved site) is seen and a band at ~120 kDa (inactive precursor) may not be seen or it may be diminished. I have added this information to the online datasheet. Our source for this antibody also provided the following information. "This product has been tested with membrane protein from normal rat liver (5 ug was enough to detect). Also, rat/mouse liver homogenate was used (50 ug was enough to detect). Normal parameters were used such as the addition of protease inhibitors and keeping the tissue as cold as possible during the protein extraction." Just recently the blocking peptide for this antibody was added to our catalog. The peptide is catalog number ab31099 and will be useful to determine which band is specific. Please contact us again if you have any additional questions.

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11-14 of 14 Abreviews or Q&A

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