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    olfm4-antibody-ab105861.pdf

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Anti-OLFM4 antibody (ab105861)

  • Datasheet
  • SDS
Reviews (2)Q&A (5)

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Western blot - Anti-OLFM4 antibody (ab105861)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-OLFM4 antibody (ab105861)

Key features and details

  • Rabbit polyclonal to OLFM4
  • Suitable for: IHC-P, WB
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-OLFM4 antibody
    See all OLFM4 primary antibodies
  • Description

    Rabbit polyclonal to OLFM4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Dog, Chimpanzee, Monkey
  • Immunogen

    Synthetic peptide corresponding to a region within internal sequence amino acids 400-450 (ASTGNMVISK LNDTTLQVLN TWYTKQYKPS ASNAFMVCGV LYATRTMNTR T) of Human OLFM4, conjugated to KLH.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • Human liver lysate
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • 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.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.05% BSA, PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cancer
    • Tumor biomarkers
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Mouse liver tissue lysate - total protein (ab29301)
  • Recombinant Protein

    • Recombinant Human OLFM4 protein (ab112393)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab105861 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
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.
WB
Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 57 kDa.
Notes
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.
WB
Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 57 kDa.

Target

  • Function

    An antiapoptotic factor that promotes tumor growth. Promotes proliferation of pancreatic cancer cells by favoring the transition from the S to G2/M phase. Facilitates cell adhesion.
  • Tissue specificity

    Expressed during myeloid lineage development. Strongly expressed in the prostate, small intestine and colon and moderately expressed in the bone marrow and stomach. Highly expressed in pancreatic cancer tissues and shows an elevated expression level during the early S phase of the cell cycle. Also expressed at high levels in stomach cancer and colon cancer tissues.
  • Sequence similarities

    Contains 1 olfactomedin-like domain.
  • Domain

    The olfactomedin-like domain is involved in the interaction with cadherin.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Secreted > extracellular space. According to PubMed:15059901, found in mitochondrion and nucleus.
  • Target information above from: UniProt accession Q6UX06 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 467367 Chimpanzee
    • Entrez Gene: 515082 Cow
    • Entrez Gene: 485470 Dog
    • Entrez Gene: 10562 Human
    • Entrez Gene: 380924 Mouse
    • Entrez Gene: 290409 Rat
    • Omim: 614061 Human
    • SwissProt: Q6UX06 Human
    • SwissProt: Q3UZZ4 Mouse
    • Unigene: 508113 Human
    • Unigene: 26456 Mouse
    • Unigene: 156265 Rat
    see all
  • Alternative names

    • Antiapoptotic protein GW112 antibody
    • bA209J19.1 antibody
    • G-CSF-stimulated clone 1 protein antibody
    • GC1 antibody
    • GW112 antibody
    • hGC 1 antibody
    • hGC-1 antibody
    • hOLfD antibody
    • KIAA4294 antibody
    • olfactomedin 4 antibody
    • Olfactomedin-4 antibody
    • OlfD antibody
    • OLFM4 antibody
    • OLFM4_HUMAN antibody
    • OLM4 antibody
    • UNQ362 antibody
    see all

Images

  • Western blot - Anti-OLFM4 antibody (ab105861)
    Western blot - Anti-OLFM4 antibody (ab105861)
    All lanes : Anti-OLFM4 antibody (ab105861) at 2 µg/ml

    Lane 1 : Human liver lysate with no immunizing peptide
    Lane 2 : Human liver lysate with immunizing peptide

    Predicted band size: 57 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-OLFM4 antibody (ab105861)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-OLFM4 antibody (ab105861)
    ab105861, at 5 µg/ml, staining OLFM4 in formalin fixed, paraffin embedded colon tissue by Immunohistochemistry. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody followed by alkaline phosphatase-streptavidin and chromogen.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab105861? Please let us know so that we can cite the reference in this datasheet.

ab105861 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Flow Cytometry abreview for Anti-OLFM4 antibody

Poor
Abreviews
Abreviews
abreview image
Application
Flow Cytometry
Sample
Human Cell (CD45+ blood cells)
Permeabilization
Yes - 1:1 methanol:acetone
Gating Strategy
Neutrophils (CD16+)
Specification
CD45+ blood cells
Preparation
Cell harvesting/tissue preparation method: Depletion of RBS from human blood using EasySep RBC depletion reagent
Sample buffer: PBS + 1% BSA
Fixation
BD Phosflow™ Lyse/Fix Buffer
Read More

Abcam user community

Verified customer

Submitted Mar 07 2020

Question

Hope you had good holidays and many thanks for your assistance. Regarding your comments: - I have tested the antibody on 2 normal cell lines (prostate and pancreas) and I still see the same strong band at ˜30kDa. As I have mentioned before I have tested the antibody on ˜20 cell lines including the 2 normal ones and all show the same band to a more or less similar extent!!!! - I have come across the paper you have mentioned. Im not sure what to make of the 1.7kb transcriptas the nature of this band has not been verified to date(the paper was published in 2004) at either transcript or protein level. Ensembl only reports 2 transcripts, 2.9kb and a non-translated 2.3kb one. The OLFM4 gene has 5 exons, exon 5 is the largest making up ˜2.1kb on its own and encodes the functional olfactomedin like domain... So Im not sure about the nature of this transcript and whether it is translated or not..... I will keep looking into that tho.... I would be grateful if you can send me the lysate and we will take it from there Regards

Read More

Abcam community

Verified customer

Asked on Jan 03 2012

Answer

Thank you for your email. I have sent the lysates ab29889 with order number 1007989. I hope this will help.

Read More

Abcam Scientific Support

Answered on Jan 03 2012

Question

Please find representative blots of both antibodies. I have tried different lysis buffers / blocking conditions/washing/antibody dilutions and the results are more or less the same. blots with only the secondary antibody are completely negative. I have so far blotted lysates from ~20 cell lines, while very few have the correct band at 57kDa, the extra bands, particularly the strong one ~30kDa are consistently present in all of them Many thanks for your help

Read More

Abcam community

Verified customer

Asked on Dec 22 2011

Answer

I have now received a reply from lab. They have confirmed observing a 57kDa using human, rat and mouse liver extracts and 57 kD/ 40 kD band in the stomach extracts. I am OK for sending positive control lysates. However I would be interested in getting your thoughts about the publication that I have sent in my previous email. Looking forward to hearing from you soon.

Read More

Abcam Scientific Support

Answered on Dec 22 2011

Question

Please find representative blots of both antibodies. I have tried different lysis buffers / blocking conditions/washing/antibody dilutions and the results are more or less the same. blots with only the secondary antibody are completely negative. I have so far blotted lysates from ~20 cell lines, while very few have the correct band at 57kDa, the extra bands, particularly the strong one ~30kDa are consistently present in all of them   Many thanks for your help

Read More

Abcam community

Verified customer

Asked on Dec 20 2011

Answer

Thank you for your email. I have forwarded the data to my colleague in lab and now I am waiting a reply. I will contact you again soon; normally they write back to us within 24 hours. I have also done further search which I would be happy in sharing with you. The 57kda molecular weight size is of unmodified protein as given in Swissprot and as per same database the OLFM4 protein is further processed in mature form with deletion of signal peptide or addition of other molecules that leads to full functioning of protein. It may well be that in the cancerous tissues or cell line OLFM4 is further processed in small isoform. Please check the attached publication (Fig1) which is supporting this; in this publication the author has observed 2 mRNA constructs in different tumour tissues size 1.7 kb and size 2.9 kb. http://cancerres.aacrjournals.org/content/64/7/2474/F1.expansion.html http://cancerres.aacrjournals.org/content/64/7/2474.long I have also carefully checked the images you have kindly provided, I am presuming that the cell lines you have used are all cancerous cell line (T3H4, HCT116, FA6 and MiaPaca2 are in fact the cancerous cell lines). Interestingly the CFPAC1 do show a band at 57KDa in addition to 37kDa. I have checked few more publications; unfortunately because cut out images are given so it was hard to understand the actual m.wt. other scientist has been observing. It will be a good of emailing few of them and discussing the results. I will contact you as soon as I receive a reply from lab. Many thanks for having patience.   

Read More

Abcam Scientific Support

Answered on Dec 20 2011

Question

Customer reported  another band at 32kD which should not be there, this even in negative samples. Does see also the correct band at 57kD. working on human samples, tried different blocking solutions, different dilutions, no primary control, samples reduced and denatured.

Read More

Abcam community

Verified customer

Asked on Dec 16 2011

Answer

Thanks you very much for contacting us. My colleagues has just forwarded your details to me. I am sorry to hear that you are observing multiple bands again. I will certainly look into this case further and would also like to discuss this case with lab. Could you be so kind sending me the image for my records? There are also 5 isoforms given in Aceview database; http://www.ncbi.nlm.nih.gov/IEB/Research/Acembly/av.cgi?db=human&term=olfm4&submit=Go. I am not sure if any of the isoform is reported in the lysates you have been using however because both antibodies gave similar results so it will be worth checking the literature. I will review the product again with data I receive from lab.  Regarding the lysates; I will send these once I finish reviewing this product. Looking forward to hearing from you soon.

Read More

Abcam Scientific Support

Answered on Dec 16 2011

Question

Can you please send me the IHC protocol used to validated ab105861?

Read More

Abcam community

Verified customer

Asked on Nov 30 2011

Answer

Thank you for contacting us.  Below is the IHC protocol for ab105861.  I hope this helps, please let me know if you have any additonal questions.   Tissue Preparation: Formalin fixation and embedding in paraffin wax Tissue Sectioning: Make 4-μm sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-drying oven for 45 minutes at 60°C. Deparaffinization: Wash dry slides in 3 changes of xylene – 5 minutes each @ RT Rehydration: Wash slides in 3 changes of 100% alcohol – 3 minutes each @ RT Wash slides in 2 changes of 95% alcohol – 3 minutes each @ RT Wash slides in 1 change of 80% alcohol – 3 minutes @ RT Rinse slides in gentle running distilled water – 5 minutes @ RT Antigen retrieval: Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes Remove from heat and let stand at room temperature in buffer - 20 minutes Rinse in 1X TBS with Tween (TBST) – 1 minute @ RT Immunostaining: (Do not allow tissues to dry at any time during the staining procedure) Apply a universal protein block – 20 minutes @ RT Drain protein block from slides, apply diluted primary antibody (5ug/mL) – 45 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply a biotinylated anti-rabbit IgG (H+L) secondary – 30 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply alkaline phosphatase streptavidin – 30 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply alkaline phosphatase chromogen substrate – 30 minutes @ RT Wash slides in distilled water – 1 minute @ RT Dehydrate:  (This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB) Wash slides in 2 changes of 80% alcohol – 1 minute each @ RT Wash slides in 2 changes of 95% alcohol – 1 minute each @ RT Wash slides in 3 changes of 100% alcohol – 1 minute each @ RT Wash slides in 3 changes of xylene – 1 minute each @ RT Apply coverslip

Read More

Abcam Scientific Support

Answered on Nov 30 2011

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-OLFM4 antibody

Inconclusive
Abreviews
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (stomach, duodenum, jejunum, ileum, colon)
Specification
stomach, duodenum, jejunum, ileum, colon
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Vector Antigen Unmasking Solution
Permeabilization
Yes - Vector Antigen Unmasking Solution, Triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C
Read More

Ms. Gail Butler

Verified customer

Submitted Aug 30 2011

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