Anti-LHX6 antibody (ab22885)

Reviews (2)Q&A (2)References (4)

Overview

  • Product name
    Anti-LHX6 antibody
    See all LHX6 primary antibodies
  • Description
    Rabbit polyclonal to LHX6
  • Host species
    Rabbit
  • Specificity
    ab22885 recognises LHX6
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat, Cow, DogDoes not react with: Mouse
  • Immunogen

    Synthetic peptide: RRVIQVWFQN CRARHKKHTP QHPVPPSGAP PSRLPSALSD DIHYTPFSSP, corresponding to a region within C terminal amino acids 289-338 of Human LHX6

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • HepG2 whole cell lysate (ab7900) and human liver tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab22885 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 Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 40 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.

Target

  • Function
    Probable transcription factor required for the expression of a subset of genes involved in interneurons migration and development. Functions in the specification of cortical interneuron subtypes and in the migration of GABAergic interneuron precursors from the subpallium to the cerebral cortex.
  • Sequence similarities
    Contains 1 homeobox DNA-binding domain.
    Contains 2 LIM zinc-binding domains.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
    • Alternative names
      • LHX 6 antibody
      • LHX 6.1 antibody
      • LHX6 antibody
      • LHX6.1 antibody
      • LHX6_HUMAN antibody
      • LIM homeobox 6 antibody
      • LIM homeobox protein 6 antibody
      • LIM homeodomain protein 6.1 antibody
      • LIM/homeobox protein Lhx6 antibody
      • LIM/homeobox protein Lhx6.1 antibody
      • MGC119542 antibody
      • MGC119544 antibody
      • MGC119545 antibody
      • OTTHUMP00000064113 antibody
      • OTTHUMP00000064114 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LHX6 antibody (ab22885)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LHX6 antibody (ab22885)
      Immunohistochemical analysis of LHX6 expression in paraffin embedded human liver tissue. ab22885 was used at 4 ug/ml. Arrows indicate positively labelled hepatocytes.
    • Western blot - Anti-LHX6 antibody (ab22885)
      Western blot - Anti-LHX6 antibody (ab22885)
      Lane 1 : Anti-LHX6 antibody (ab22885) at 5 µg/ml
      Lane 2 : Anti-LHX6 antibody (ab22885) at 2.5 µg/ml
      Lane 3 : Anti-LHX6 antibody (ab22885) at 1.25 µg/ml

      All lanes : Cell lysate prepared from human HepG2 cells

      Lysates/proteins at 25 µg per lane.

      Predicted band size: 40 kDa
      Observed band size: 50 kDa
      why is the actual band size different from the predicted?

    References

    This product has been referenced in:
    • Maroof AM  et al. Directed differentiation and functional maturation of cortical interneurons from human embryonic stem cells. Cell Stem Cell 12:559-72 (2013). Read more (PubMed: 23642365) »
    • García-Moreno F  et al. A neuronal migratory pathway crossing from diencephalon to telencephalon populates amygdala nuclei. Nat Neurosci 13:680-9 (2010). IHC-P ; Mouse . Read more (PubMed: 20495559) »
    See all 4 Publications for this product

    Publishing research using ab22885? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-LHX6 antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human fibroblast and stem cells)
    Loading amount
    50 µg
    Specification
    human fibroblast and stem cells
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 10 2012

    Question

    BATCH NUMBER 222072 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Ab identifies MOUSE Lhx6 (aprox. 50KDa band). However, in 293T cells (HUMAN), there are additional bands STRONGER than the 50KDa band and of HIGHER MW (degradation should be excluded) SAMPLE HEK 293T cells transfected with GFP (control) or GFP+ mouseLHX6 (vector pCAGGs). Whole cell extract PRIMARY ANTIBODY Abcam Lhx6 polyclonal antibody (ab22885) 1:2,000 in 2.5%milk in PBST o/n to 48h, 4oC (does not make any difference. Wash in PBST 5x10', RT. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED GFP TRANSFECTED (NEGATIVE) vs MOUSE LHX6 TRANSFECTED CELLS EXPRESSION OF PROTEIN TESTED ON IHC USING TWO DIFFERENT LHX6 Abs (Abcam and another polyclonal ANTIBODY STORAGE CONDITIONS -20oC in aliquots. Once defrosted, stored at 4oC. SAMPLE PREPARATION Lysis buffer: 50 mM TRIS pH7.5, 150 mM NaCl, 0.1%SDS, 1%NP-40, 0.5% Nadeoxycholate + EDTA free protease inhibitors. AMOUNT OF PROTEIN LOADED Not quantitated (however, ACTIN control immunodetection shows equivalent amounts in all lanes. ONE well (6-well plate) is lyzed in 500 ul LYSIS buffer and 10 ul run on gel ELECTROPHORESIS/GEL CONDITIONS 10% gel SDS-PAGE using standard protocol (Current Protocols) TRANSFER AND BLOCKING CONDITIONS Semidry transfer using standard protocol and buffer (as recommended by company) Block in 5% milk in PBST (0.1%Tween20 in PBS,)4h to o/n 4oC (does not make any difference). SECONDARY ANTIBODY GOAT ANTI RABBIT IgG(H+L) HRP CONJUGATE (BIORAD Cat.No. 172-1013) incubate 2h, RT Wash 5X10', RT in PBST HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? TRIED TO HOMOGENIZE,SOLUBILIZE CELL LYSATE BETTER. ADDITIONAL NOTES since my problem is additional bands in a specific cell line, my question is whether you have tested the ab in a different cell line other than HepG2 and of course I want to know whether you used a similar protocol to the above.

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    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. Thank you also for the image that you have provided me with, it is most useful in understanding the difficulties that you have experienced. As I understand you have been detecting very strong bands in cells transfected with GFP fused to murine LHX6 in addition to cells transfected with GFP alone. For the testing of this antibody we used HepG2 cell lysates and obtained a single band at ~55KDa. I would like to ask whether the intense band that you have been detecting between 50-75KDa could possibly be this product? Do you have an image of this blot with a reduced exposure? Does it form a clean band? We do see variation in the detectable molecular weight dependent on the set of molecular markers employed. I appreciate that this does not explain the absence of detection of the murine Lhx6-GFP construct. However, this antibody has to date not yet shown to have reactivity against the mouse homolog and may explain the results that you have been obtaining. I appreciate your input into this.

    Read More

    Question

    DESCRIPTION OF THE PROBLEM The Ab recognized three major bands (of 105, 60 and 35 kDa) and a minor one of Mr 50kDa SAMPLE mouse brain PRIMARY ANTIBODY Washed with TPBS and incubation with Rabbit polyclonal antibody antiLhx6 Ab22885 1ug/ml in TPBS over night at -4ºC DETECTION METHOD ECL plus POSITIVE AND NEGATIVE CONTROLS USED none ANTIBODY STORAGE CONDITIONS storage at -40ºC SAMPLE PREPARATION total proteins extracted with 4% SDS plus 8M urea AMOUNT OF PROTEIN LOADED 10 ug ELECTROPHORESIS/GEL CONDITIONS polyacrylamide gel 12% TRANSFER AND BLOCKING CONDITIONS blocking with milk/TPBS 5% (1 hour) SECONDARY ANTIBODY Wash with TPBS and incubation with HRP polyclonal antirabbit 1/6000 in TPBS for 1 hour at RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No

    Read More
    Answer

    Thank you for your enquiry. I am sorry to hear you are having a problem with ab22885 (LHX6 antibody). To improve the detection of LHX6, I would like to suggest the following modifications to your protocol: 1) As LHX6 is a nuclear protein, use RIPA buffer to lyse your cells and load more protein samples e.g. 50-100 ug. This might increase the signal of LHX6 but also unspecific bands. 2) Include a "secondary antibody control" (no ab22885) and adjust the dilution of the 2nd antibody accordingly (e.g. dilute 1/10,000). 3) Increase blocking time to 2hr room temperature and/or try another blocking reagent e.g. BSA. I want to point out that ab22885 has not been tested on mouse LHX6 therefore we can not guarantee that it will work on this species. Your results actually suggest that ab22885 might recognize mouse LHX6 but possibly also other mouse proteins. Please let me know if this improves your results. I look forward to your reply.

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    Immunohistochemistry (Frozen sections) abreview for Anti-LHX6 antibody

     Inconclusive
    Abreviews
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (brain, fixed in acrolein)
    Specification
    brain, fixed in acrolein
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 6%
    Read More

    Miss. Nina Hasen

    Verified customer

    Submitted Jun 01 2006

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