Overview

  • Product name
  • Description
    Goat polyclonal to Bim
  • Host species
    Goat
  • Specificity
    This antibody is expected to recognise 3 reported isoforms: BimAD, BimACD, and BimABCD
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide:

    FNAYYARRLEK

    , corresponding to amino acids 159-169 of Human Bim.

Properties

Applications

Our Abpromise guarantee covers the use of ab17031 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 0.5 - 2 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 19, 9, 12 kDa).

Target

  • Function
    Induces apoptosis. Isoform BimL is more potent than isoform BimEL. Isoform Bim-alpha1, isoform Bim-alpha2 and isoform Bim-alpha3 induce apoptosis, although less potent than the isoforms BimEL, BimL and BimS. Isoform Bim-gamma induces apoptosis.
  • Tissue specificity
    Isoform BimEL, isoform BimL and isoform BimS are the predominant isoforms and are ubiquitously expressed with a tissue-specific variation. Isoform Bim-gamma is most abundantly expressed in small intestine and colon, and in lower levels in spleen, prostate, testis, heart, liver and kidney.
  • Sequence similarities
    Belongs to the Bcl-2 family.
  • Domain
    The BH3 motif is required for Bcl-2 binding and cytotoxicity.
  • Cellular localization
    Mitochondrion and Endomembrane system. Associated with intracytoplasmic membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • BCL2 like 11 antibody
    • B2L11_HUMAN antibody
    • BAM antibody
    • Bcl 2 interacting protein Bim antibody
    • Bcl 2 related ovarian death agonist antibody
    • Bcl-2-like protein 11 antibody
    • BCL2 interacting mediator of cell death antibody
    • BCL2 like 11 (apoptosis facilitator) antibody
    • BCL2 like protein 11 antibody
    • Bcl2-interacting mediator of cell death antibody
    • Bcl2-L-11 antibody
    • Bcl2l11 antibody
    • BIM alpha6 antibody
    • BIM antibody
    • BIM beta6 antibody
    • BIM beta7 antibody
    • BimEL antibody
    • BimL antibody
    • BOD antibody
    see all

Images

  • Anti-Bim antibody (ab17031) at 0.5 µg/ml + Cell lysates prepared from Human K562 cells at 35 µg

    Predicted band size: 19, 9, 12 kDa



    ab17031 at 0.5ug/ml on K562 lysate (35ug).

References

This product has been referenced in:
  • Holmberg C  et al. Release of TGFßig-h3 by gastric myofibroblasts slows tumor growth and is decreased with cancer progression. Carcinogenesis 33:1553-62 (2012). Human . Read more (PubMed: 22610072) »
See 1 Publication for this product

Customer reviews and Q&As

Question


Thanks for your quick feedback, attached you will find images of my WB experiment. The questionnaire is below this messege.
Best regards





1) Abcam product code ab 17030


2) Abcam order reference number or product batch number: Order number (290_PiNi_0124) Lot: GR 74237-1


3) Description of the problem

Failure to obtain positive results (the right band was not observed)

4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysate

Lysis buffer : *competitor* ready made buffer
Protease inhibitors- Yes
Phosphatase inhibitors: Yes
Reducing agent: Yes
Boiling for ≥5 min? yes (10 min)
Protein loaded ug/lane or cells/lane: (30ug)
Positive control : None
Negative control: ?

5) Percentage of gel :12%
Type of membrane : PVDF and Nitro
Protein transfer verified : Yes by staining the membrane and the gel after transfere
Blocking agent and concentration: 3% BSA or 2.5% milk powder
Blocking time: Overnight at 4ºC and 30 min at RT
Blocking temperature

6) Primary antibody (If more than one was used, describe in “additional notes”)
Concentration or dilution 1:1000 and 1:500
Diluent buffer : 3% BSA or 2.5% milk powder
Incubation time: 1h or overnight
Incubation temperature: RT or 4 ºC

7) Secondary antibody:
Species:
Reacts against: Goat
Concentration or dilution 1:30000
Diluent buffer: 3% BSA or 2.5% milk powder
Incubation time:1h
Incubation temperature: RT
Fluorochrome or enzyme conjugate: HRP

8) Washing after primary and secondary antibodies:
Buffer: TBST and TBST+Triton
Number of washes X3

9)Detection method : ECL
10) How many times have you run this staining : 3X
Do you obtain the same results every time? NO
What steps have you altered to try and optimize the use of this antibody? Blcoking, Buffers,antibody concentration.

Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab17031.

1.) I suggest to review the lysis buffer that is used. I found the following information: *competitor* ready made buffer can solubilize membrane proteins as well as cytoplasmic proteins and the conditions are non-denaturing. However, we cannot guarantee the solubilization of all membrane proteins.

Bim is a membrane protein on the endomembanre system and mitochondria. maybe this buffer is not optimal. I can recommend to use a standardRIPA buffer (RIPA buffer: 20mM Tris-HCL pH7.4, 150mM Na\Cl, 1mM EDTA, 1% Triton-X100, 1% sodium deoxycholate, 0.1% SDS with freshly added PMSF to 1mM and with freshly added aprotinin and leupeptin to 5ug/ml just before use.)

2.) I can also recommend to test a different cell line. Bim is expressed ubiquitously and this will then show if there is a specific problem with the cells you are using.

3.) Since the protein is very small, I suggest to make sure it did not run out of the gel. Without molecular weight marker information, I cannot judge this from the image.

I would also appreciate if you can confirm some further details:

1.) Did the loading control work?

2.) Was the secondary tested with a different primary?

3.) What cells were used? Tissue and species?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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