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Anti-TNF alpha antibody [52B83] (ab1793)

Reviews (18)Q&A (33)References (238)
Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [52B83] (ab1793)
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

Key features and details

  • Mouse monoclonal [52B83] to TNF alpha
  • Suitable for: ELISA, IHC-P, ICC/IF, Flow Cyt (Intra), IHC-Fr, WB
  • Reacts with: Mouse, Guinea pig, Human, Chimpanzee, Zebrafish, Cynomolgus monkey, Rhesus monkey, Apteronotus leptorhynchus
  • Isotype: IgG1

Overview

  • Product name

    Anti-TNF alpha antibody [52B83]
    See all TNF alpha primary antibodies

  • Description

    Mouse monoclonal [52B83] to TNF alpha

  • Host species

    Mouse

  • Specificity

    This antibody detects both natural and recombinant TNFa. It does not cross-react with TNF beta or lymphotoxin. It reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. It does not react with receptor-bound TNF-alpha. Non-specific background staining is observed in connective tissues.

  • Tested applications

    Suitable for: ELISA, IHC-P, ICC/IF, Flow Cyt (Intra), IHC-Fr, WBmore details

  • Species reactivity

    Reacts with: Mouse, Guinea pig, Human, Chimpanzee, Zebrafish, Cynomolgus monkey, Rhesus monkey, Apteronotus leptorhynchus

  • Immunogen

    Full length native protein (purified) corresponding to Human TNF alpha.

  • Positive control

    • Flow Cyt (Intra): RAW 264.7 cells.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab1793 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
ELISA
Use at an assay dependent concentration.
IHC-P (5)
Use a concentration of 2.5 µg/ml.
ICC/IF (2)
Use a concentration of 5 µg/ml.
Flow Cyt (Intra)
1/10.

(methanol fixed cells)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr (3)
Use a concentration of 2 µg/ml.

Fixed in acetone for 10 minutes
WB (8)
Use a concentration of 2 µg/ml. Predicted molecular weight: 25 kDa.

TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha is 17.4 kDa A reduced sample treatment and 15% SDS-Page was used.
Notes
ELISA
Use at an assay dependent concentration.
IHC-P
Use a concentration of 2.5 µg/ml.
ICC/IF
Use a concentration of 5 µg/ml.
Flow Cyt (Intra)
1/10.

(methanol fixed cells)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr
Use a concentration of 2 µg/ml.

Fixed in acetone for 10 minutes
WB
Use a concentration of 2 µg/ml. Predicted molecular weight: 25 kDa.

TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha is 17.4 kDa A reduced sample treatment and 15% SDS-Page was used.

Target

  • Function

    Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation.

  • Involvement in disease

    Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).

  • Sequence similarities

    Belongs to the tumor necrosis factor family.

  • Post-translational
    modifications

    The soluble form derives from the membrane form by proteolytic processing.
    The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
    O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.

  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • APC1 antibody
    • APC1 protein antibody
    • Cachectin antibody
    • DIF antibody
    • Differentiation inducing factor antibody
    • Macrophage cytotoxic factor antibody
    • Tnf antibody
    • TNF superfamily member 2 antibody
    • TNF superfamily, member 2 antibody
    • TNF, macrophage derived antibody
    • TNF, monocyte derived antibody
    • TNF-a antibody
    • TNF-alpha antibody
    • TNFA antibody
    • TNFA_HUMAN antibody
    • TNFSF2 antibody
    • Tumor necrosis factor (TNF superfamily member 2) antibody
    • Tumor necrosis factor alpha antibody
    • Tumor necrosis factor antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor Necrosis Factor, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    ab1793 staining TNFα in RAW 264.7 cells treated with (R,S)-rolipram (ab120029), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R,S)-rolipram, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120029 ((R,S)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [52B83] (ab1793)
    Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [52B83] (ab1793)
    Overlay histogram showing RAW 264.7 cells stained with ab1793 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1793, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in RAW 264.7 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    ab1793 staining TNFα in RAW 264.7 cells treated with (R)-(-)-rolipram (ab120031), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R)-(-)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120031 ((R)-(-)--rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

    ab1793 staining TNFα in RAW 264.7 cells treated with (S)-(+)-rolipram (ab120030), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (S)-(+)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120030 ((S)-(+)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References (238)

Publishing research using ab1793? Please let us know so that we can cite the reference in this datasheet.

ab1793 has been referenced in 238 publications.

  • Zheng Y  et al. Low-level laser therapy prevents medication-related osteonecrosis of the jaw-like lesions via IL-1RA-mediated primary gingival wound healing. BMC Oral Health 23:14 (2023). PubMed: 36627695
  • Zhou T  et al. miR-204-containing exosomes ameliorate GVHD-associated dry eye disease. Sci Adv 8:eabj9617 (2022). PubMed: 35020440
  • Park HR & Yang EJ Combined Treatment with Herbal Medicine and Drug Ameliorates Inflammation and Metabolic Abnormalities in the Liver of an Amyotrophic Lateral Sclerosis Mouse Model. Antioxidants (Basel) 11:N/A (2022). PubMed: 35052677
  • Du J Synergistic effect of Bruton's tyrosine kinase and TNF-α in the regulation of rheumatoid arthritis and underlying mechanisms. Exp Ther Med 23:141 (2022). PubMed: 35069822
  • Hu M  et al. Effect of α-tocopherol in alleviating the lipopolysaccharide-induced acute lung injury via inhibiting nuclear factor kappa-B signaling pathways. Bioengineered 13:3958-3968 (2022). PubMed: 35112986
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

21-30 of 51 Abreviews or Q&A

Question

Thank you for the help but I do havea few more questions.
On the websiteI saw a abreview submitted on 19 July 2010where they claim that the specific band of TNF-alpha in zebrafishis around 40 kDa. ShouldIbelieve on this or would it be rather around 60 kDa?
Can I alsouse alpha-tubulin (ab15246) asa loadingcontrol for zebrafish? I rather would like to use this one because I saw that some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types

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Answer

Thank you for your reply.
Unforunately, I do not have any information about what the exact zebrafish band size should be for TNF-alpha. Based on the UniProt database, the predicted molecular weight for zebrafish is 25.8 kDa: http://www.uniprot.org/uniprot/Q1L8A5. The UniProt database also provides information about the presence of a secreted homotrimer in human (the human protein is more thoroughly characterized than the zebrafish protein): http://www.uniprot.org/uniprot/P01375.
It is definitely possible that the Abreviewer may be aware of literature that would indicate the zebrafish band should have an apparently molecular weight of 40 kDa. While this Abreview was submitted anonymously, the reviewer has selected to enable messages, so I would recommend contacting him or her directly for additional information about those results. This can be done by clicking the link at the top of the Abreview.
Alpha-Tubulin can also be used as a loading control, and ab15246 will be guaranteed for use on zebrafish samples. You may also be interested in the following flyer that lists our current range of antibodies validated for zebrafish:
https://docs.abcam.com/pdf/general/Zebrafish_Antibodies_with_cross_reactivity.pdf
I hope this helps, please let me know if you need any additional information or assistance.

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Question

LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands around 80kDa, 60kDa and 40 kDa. I would like to know the specifity of the anti-TNF alpha Ab for zebrafish. Do you have peptides to block the antibody binding? SAMPLE Danio Rerio (zebrafish), cell extract PRIMARY ANTIBODY ab1793 1/1000 overnight incubation DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED i would like to use beta-actin as positive control, do you have an antibody for zebrafish? SAMPLE PREPARATION cell extraction buffer (invitrogen), PMSF and complete (Roche) AMOUNT OF PROTEIN LOADED 40 µg TRANSFER AND BLOCKING CONDITIONS 10xTBS, milk,Tween. SECONDARY ANTIBODY secondary mouse antibody 1/2000 2.5 h incubation

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Answer

Thank you for contacting us. I am sorry that this antibody is producing multiple bands on zebrafish samples.
We do not have a blocking peptide available for use with ab1793 because it was raised against the full length native human TNF alpha protein. While none of our beta actin antibodies have been validated for use on zebrafish, our GAPDH antibody ab8245 has. GAPDH is a similar cytoplasmic loading control and ab8245 would be guaranteed for your applications.
The predicted molecular weight of TNF alpha is 17.4 kDa, however, this protein is normally secreted as a homotrimer with a molecular weight of 52 kDa. It is likely that the band you are seeing around 60 kDa is the secreted form of TNF alpha. To lower the intensity of the other bands, you could try reducing the concentration of your secondary antibody to 1:5000 - 1:10,000. It may also help to block for a longer amount of time with 5% milk.
I hope this helps, if not, please let me know your original order number and I will be happy to help you further.

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Question

I am thinking about ordering ab6672 and ab1793. However I'm a bit worried that the antibodies may detect non-specific bands as I've never used them before. I am wondering whether you could include the positive control for each antibody that I order. The positive control will at least give me peace of mind that the antibodies detect the right bands. I understand that it might not be possible to obtain a small sample of positive control at no extra cost, but I thought I give it a go. Hope you could help us out.

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Answer

Thank you for contacting us.

Unfortunately we do not provide positive controls with our antibodies.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Question

HI

I NEED CERTIFICATE OF ANALYSIS OF THE FOLLOWING PRODUCTS:

CAT. NUMBER LOT

AB27969 GR66929-1

AB1793 GR66247-1

AB2105 GR24376-3

AB6672 GR58964-2

AB34843 GR32935-5

AB97049 GR46176-1

THANKS

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Answer

Thank you for contacting Abcam.

Please find attached the requested Certificate of Compliances.

If there is anything else I can help you, please let me know.

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Question

Your answer gave me no solution for my problem. Unfortunately. I think your suggestion to give me a credit note is the best solution. Please send me a credit note.
I will include my order form of the Abcam ab1793.

We have an answer to your inquiry:
Thank you for providing the additional information and western blot data. That was very helpful.
I agree that the western blot data usingab1793 does notproduce a band at the expected molecular weight andif you havereceived this antibody within the last 6 months or so I would be happy to offer a replacement, creditor refund.
Regarding, ab66579, it seems that the rat liver supernatant produces a band at the expected molecular weight and at a mass comparable to the western blot data shown on the online datasheet for ab66579 using a 1:500 dilution.
Please let me know what purchase order number or Abcam order reference number was used to purchase the vial of ab1793 so I can process your request.

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Answer

I am glad you followed up with me as I have searched our records and unfortunately we did not receive the email you sent on 12/5/2011.

I also apologize that my reply was not helpful for you. I would be happy to hear of any suggestions to help me provide you with more useful information.

I am currently awaiting approval from our accounts teamin order to issue the credit note for ab1793. As soon as I do I will send you the credit note number and details on how to redeem the note.

Thanks for your patience!

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Question

DESCRIPTION OF THE PROBLEM The background is too dark and the wrong bands ( not shown in the matrix)
SAMPLE Mouse Tissue
PRIMARY ANTIBODY 1:100 0r 1:200 dilution 37°C incubate 1 hour wash buffer:PBS wash 3 times 15min/time
DETECTION METHOD DAB
POSITIVE AND NEGATIVE CONTROLS USED no
ANTIBODY STORAGE CONDITIONS Store at +4°C
FIXATION OF SAMPLE Formalin
ANTIGEN RETRIEVAL Yes, it does. Heat treatment by pressure cooker or microwave oven. Also, we have tried the enzyme digestion but it still does not work.
PERMEABILIZATION STEP no
BLOCKING CONDITIONS Species of serpurn exactly goat
SECONDARY ANTIBODY rabbit anti-mouse.
HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2
HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes
DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
WHAT STEPS HAVE YOU ALTERED? different dilution of primary Antibody

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Answer

Thank you for contacting us regarding this issue.
I was hoping that you might clarify a few points about the staining procedure you use with this product. In particular I was wondering the formulation of your blocking solution, how long and at what temperature was your blocking step performed? I have also noticed that the recommended dilution for this antibody is 1:50, have you tried this dilution or one which is greater, perhaps 1:25?
I look forward to your reply. Please do not hesitate to contact us if you need any more advice or information.

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Question

Hello, another scientific support specialist helped us previously with a testing offer for the TNF alpha antibody (ab6671) in squirrel. I have finished testing the antibody in our squirrel samples and am ready to submit an abreview. I have a question however. Do I have to submit the review under the same account used to make the purchase. Orders from our lab are made using the account of our lab manager and expediter and so we would like to apply the promotional code, to our next order. Before I submit my review I would like to know if it should be submitted using our expediters account. The same question would apply to our other post-docs who have antibodies they can review but want to make sure we can use the acquired abpoints collectively for our lab. Also, I would like ask for a promotional code for testing ab109555 in the squirrels. There are two differences in amino acid sequence from the immunogen but they are conserved substitutions which makes it as good a match as the rat alignment; thus, we think there is a reasonable chance this one will work for the squirrel. I have one last question. Is there a process to critique some of the submitted abreviews? While the information is potentially invaluable, I have come across some reviews which raise more doubt than support for a given product. For example, ab1793 has a review for WB with this antibody in zebrafish although the submitted image seems to contradict the "good" rating in this species. Thank you very much. As soon as I hear back on submitting the abreview I will get it posted.

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Answer

Thank you for contacting us with these questions. First, I'd like to let you know that you do not have to submit the review under the same account that ordered as your purchaser. The rewards may be used collectively. Secondly, there is no way at present to critique submitted abreviews. Finally, I'd like to offer you a 100% Abreview discount for using ab109555 in squirrel. DISCOUNT CODE: xxxx Expiration date: xxxx-xx-xx I am very pleased to hear you would like to accept our offer and test ab109555 in squirrel. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for squirrel and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question

ab1793: IHC-P image legend says "ab1793 staining TNF alpha in mouse liver tissue sections ... An undiluted HRP-conjugated monoclonal to rabbit IgG (1/200) was used as secondary antibody. " But the primary antibody is a mouse monoclonal Ab. will test ab687 in mouse (homology is 74%)

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Answer

Thank you for your phone call. 1) Regarding the image for ab1793, this was submitted by a customer as Abreview. For the details of the Abreview please follow this link: https://www.abcam.com/TNF-alpha-antibody-52B83-ab1793.html&tab=abreviews&intabreviewid=17818 It might well be that the monoclonal rabbit IgG secondary antibody is part of the DAKO EnVision+ System, Peroxidase/DAB kit mentioned in the Abreview. But unfortunately, I do not have more information in this regard. ------------------------------------------------------------------------- 2) As for the testing discount for ab687, here is the necessary information: DISCOUNT CODE: xxx Expiration date: xxx I am very pleased to hear you would like to accept our offer and test ab687 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. I hope this information is of help to you. Please let me know if I can assist you further with our products.

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Question

Hi,    thank you for your interaction. I get a signal after blocking in BSA 5% after 15 min film exposition. but, I'm not convinced about the molecular weight of TNF alpha I got. its close to 51kda!! I was expecting the band at 17kda!! the gel was under denaturing conditions (boiled samples) and loading buffer was with reducing agent DTT. thank you nabila  

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Answer

TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha (17.4 kDa) is not biologically active. Therefore what you are seeing maybe correct. To try and break these complexes down further, I would recommend increasing your boiling time to 10 minutes and that should break down these complexes. Please let me know if there is anything else I can help you wi

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Question

 Hi abcam,  Thank you for the suggestions for ab 1793.  tried to incubate again  the same filter at 1:500 overnight 4°c. I dont have signal. The filter was blocked first time in milk. I checked the paper of Mastronardi FG et al., 2006 , in which the same antibody has been used, mentioned in the abcam website. in this paper, they used milk 5% for antibodies incubation! Im wondering either I could block again the same filter in BSA and re-incubate antibody in BSA too. or I need to run a new gel... thank you

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Answer

Thank you for your reply. I think it would be OK to go ahead and reprobe the same membrane. I would recommend washing the membrane in PBS and then blocking it in 5%BSA and incubate that O/N at 4C. Please let me know if this proves to be successful or not.  

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21-30 of 51 Abreviews or Q&A

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