Overview

  • Product name
  • Description
    Rabbit polyclonal to Galectin 7
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chimpanzee, Gorilla, Orangutan
  • Immunogen

    Immunogen was a synthetic peptide, which represented a portion of human Lectin, Galactoside-binding 7 encoded within exon 4 (LocusLink ID 3963).

  • Positive control
    • Modified RIPA extract from the epidermis of FVB wild-type or galectin 7 transgenic mice.
  • General notes


    The galectins are a family of beta-galactoside-binding proteins implicated in modulating cell-cell and cell-matrix interactions. Differential and in situ hybridizations indicate that this lectin is specifically expressed in keratinocytes. It is expressed at all stages of epidermal differentiation (i.e., in basal and suprabasal layers). The protein was found mainly in stratified squamous epithelium. It is moderately repressed by retinoic acid. The antigen localized to basal keratinocytes, although it was also found at lower levels in the suprabasal layers, where it concentrated to areas of cell-to-cell contact. The cellular localization and its striking down-regulation in cultured keratinocytes imply a role in cell-cell and/or cell-matrix interactions necessary for normal growth control.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antibodies were affinity purified using the peptide immobilized on solid support.
  • Primary antibody notes
    The galectins are a family of beta-galactoside-binding proteins implicated in modulating cell-cell and cell-matrix interactions. Differential and in situ hybridizations indicate that this lectin is specifically expressed in keratinocytes. It is expressed at all stages of epidermal differentiation (i.e., in basal and suprabasal layers). The protein was found mainly in stratified squamous epithelium. It is moderately repressed by retinoic acid. The antigen localized to basal keratinocytes, although it was also found at lower levels in the suprabasal layers, where it concentrated to areas of cell-to-cell contact. The cellular localization and its striking down-regulation in cultured keratinocytes imply a role in cell-cell and/or cell-matrix interactions necessary for normal growth control.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10482 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 1/10000 - 1/40000. Detects a band of approximately 14 kDa (predicted molecular weight: 16 kDa).
IP Use at 10-20 µg/mg of lysate.
ICC/IF 1/1000.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Could be involved in cell-cell and/or cell-matrix interactions necessary for normal growth control. Pro-apoptotic protein that functions intracellularly upstream of JNK activation and cytochrome c release.
  • Tissue specificity
    Mainly expressed in stratified squamous epithelium.
  • Sequence similarities
    Contains 1 galectin domain.
  • Cellular localization
    Cytoplasm. Nucleus. Secreted. May be secreted by a non-classical secretory pathway.
  • Information by UniProt
  • Database links
  • Alternative names
    • Gal-7 antibody
    • GAL7 antibody
    • Galectin 7B antibody
    • Galectin-7 antibody
    • HKL-14 antibody
    • Human keratinocyte lectin 14 antibody
    • Keratinocyte lectin 14 antibody
    • Lectin galactoside binding soluble 7 antibody
    • Lectin, galactoside binding, soluble, 7B antibody
    • LEG7_HUMAN antibody
    • LGALS7 antibody
    • LGALS7A antibody
    • LGALS7B antibody
    • P53 induced protein 1 antibody
    • p53-induced gene 1 protein antibody
    • Pi7 antibody
    • PIG1 antibody
    • TP53I1 antibody
    see all

Images

  • ab10481 detection of mouse galectin 7 by Western blot and immunoprecipitation. Samples: A. Modified RIPA extract (10 µg) from the epidermis of FVB wild-type (wt) or galectin 7 transgenic (TG) mice. B. Modified RIPA extract (20 µg) from the epidermis of FVB wild-type mice. Antibody: ab10482 used at 0.005 (A) or 0.0025 (B) µg/ml for WB and as indicated for IP. Detection: Chemiluminescence with 1 second exposure time.

    ab10481 detection of mouse galectin 7 by Western blot and immunoprecipitation. Samples: A. Modified RIPA extract (10 µg) from the epidermis of FVB wild-type (wt) or galectin 7 transgenic (TG) mice. B. Modified RIPA extract (20 µg) from the epidermis of FVB wild-type mice. Antibody: ab10482 used at 0.005 (A) or 0.0025 (B) µg/ml for WB and as indicated for
  • ab10482 staining Galectin7 in Human cervical tumor and immortalized keratinocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100/PBS and blocked with 1% BSA for 1 hourat 25°C. Samples were incubated with primary antibody (1/1000 in 1% BSA/PBS) for 24 hours at 4°C. An Alexa Fluor®594-conjugated Goat anti-rabbit IgG polyclonal (1/2000) was used as the secondary antibody.
    SiHa cells (negative control - very low Gal-7 mRNA)
    Immortilized keratinocytes (positive control - high Gal-7 mRNA)
  • IHC image of ab10482 staining in human Hodgkin's lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10482, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:
  • Lionnard L  et al. TRIM17 and TRIM28 antagonistically regulate the ubiquitination and anti-apoptotic activity of BCL2A1. Cell Death Differ N/A:N/A (2018). Read more (PubMed: 30042493) »
  • Advedissian T  et al. E-cadherin dynamics is regulated by galectin-7 at epithelial cell surface. Sci Rep 7:17086 (2017). Read more (PubMed: 29213102) »
See all 6 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Keratinocytes)
Specification
Keratinocytes
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% triton-x-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Keratinocytes)
Loading amount
20 µg
Specification
Keratinocytes
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (schwannoma cell lines, LAN1, U87)
Loading amount
75 µg
Specification
schwannoma cell lines, LAN1, U87
Gel Running Conditions
Reduced Denaturing (12.5-15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (cervical tumor and immortalized keratinocytes)
Specification
cervical tumor and immortalized keratinocytes
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton X-100 / PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 17 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (keratinocytes and cervical tumors)
Loading amount
80 µg
Specification
keratinocytes and cervical tumors
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Bladimiro Rincon Orozco

Verified customer

Submitted Jun 30 2010

Answer

Thank you for your enquiry regarding ab10482 and for providing some further information. I am very sorry to hear that you are having some problem with this antibody. I understand from your e-mail that you used lymfoblast cell lysate. Can you please specify the species? Blocking is very important step and the time and the temperature is crucial. Normally I suggest blocking the membrane at least for 1 hr at 4oC – it seems that 10 min is too short. For further information on blocking and Western blot, please visit our website at: https://www.abcam.com/index.html?pageconfig=resource&rid=10413 https://www.abcam.com/assets/pdf/protocols/WB-beginner.pdf Have you tried to use 5% BSA for blocking instead of milk? Milk sometimes can produce non-specific bands on the blot and in our lab we usually use BSA for blocking. I would also recommend testing the secondary antibody alone i.e. run a no primary control to see if the non-specific bands are due to the secondary antibody or not. I hope this will help. Should you still have problem with this antibody, then please do not hesitate to contact me again.

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Question

BATCH NUMBER 94579 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM We detected a weak band near 17 kDa, and many bands (250~37kDa, main bands were about 37k, 50K, 60k bands). Please give me the information about preparation method of positive control (Modified RIPA extract from the epidermis of mice, and so on). SAMPLE adult mouse spinal cord homogenate (According to analysis of mass spectrometry, this sample contains galectin 7. ) PRIMARY ANTIBODY Dilution: 1/1000 in Blocking buffer Incubation time; 1 hr Wash step; 5 min, 5 min, 10 min (T-TBS) DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED negative control; no primary control ANTIBODY STORAGE CONDITIONS 4 degrees C storage SAMPLE PREPARATION Denature using SDS; incubation at RT Homogenization Buffer; 0.32 M sucrose, 50 mM Tris-HCl, pH 7.5, 1 mM EDTA, 1 mM sodium orthovanadate, 20 mM sodium diphosphate decahydrate, 20 mM glycerophosphate, 1 mM DTT, 0.75 um aprotinin, 1 mM leupeptin, 1 mM pepstatin, 0.4 mM phenylmethylsulfonyl fluoride, pH 7.5. AMOUNT OF PROTEIN LOADED 20 micrograms ELECTROPHORESIS/GEL CONDITIONS Non-reducing 13.5% gel TRANSFER AND BLOCKING CONDITIONS Transfer Buffer; 25 mM Tris base, 192 mM glycine containing 10% methanol Time; 30 V, 1000 min. Blocking agent; skimmilk / 0.1% Tween 20, 0.15 M Nacl, 20 mM Tris-HCl,pH 7.4 (T-TBS) SECONDARY ANTIBODY Santa Cruz, goat anti-rabbit IgG antibody Dilution: 1/2000 in T-TBS Incubation time; 30 min Wash step; 5 min, 5 min, 10 min (T-TBS) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes

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Answer

Thank you for your enquiry and your patience. Here we are forwarding the protocol for Preparation of Epidermal Lysates: Transgenic and nontransgenic mice were killed by cervical dislocation. The dorsal skins were treated with a depilatory agent (1 min) followed by washing. The skin was excised, and the epidermis was scraped off with a razor blade into lysis buffer [50 mm Tris (pH 7.4), 1% Triton X-100, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 1 mM Na3VO4, 1 mM NaF, and 1 ug/ml each of aprotinin, leupeptin, and pepstatin]. The lysates were homogenized using a needle (18-gauge) and syringe and were centrifuged at 14,000 x g for 15 min at 4°C. The supernatant was used immediately for Western blot analysis. We hope this information will be useful for you. If you need anything further or any help then please let us know.

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