Anti-LC3A/B antibody (ab128025)

Rabbit polyclonal LC3A/B antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Cow, Human, Drosophila melanogaster, Non human primates. Cited in 23 publication(s).

Reviews (8)Q&A (1)References (35)

Overview

  • Product name
    Anti-LC3A/B antibody
    See all LC3A/B primary antibodies
  • Description
    Rabbit polyclonal to LC3A/B
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Drosophila melanogaster, Non human primates
  • Immunogen

    Synthetic peptide corresponding to Human LC3A/B (C terminal).
    Database link: Q9GZQ8

  • Positive control
    • HeLa whole cell lysates Human Cerebral Cortex Mouse ES cell lysate

Applications

Our Abpromise guarantee covers the use of ab128025 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
ICC/IF 1/100.
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 15 kDa.
IHC-P Use a concentration of 5 - 10 µg/ml.

Target

  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • ATG8F antibody
    • Autophagy related protein LC3 A antibody
    • Autophagy related protein LC3 B antibody
    • Autophagy related ubiquitin like modifier LC3 A antibody
    • Autophagy related ubiquitin like modifier LC3 B antibody
    • Autophagy-related protein LC3 B antibody
    • Autophagy-related ubiquitin-like modifier LC3 B antibody
    • LC3 antibody
    • MAP1 light chain 3 like protein 1 antibody
    • MAP1 light chain 3 like protein 2 antibody
    • MAP1 light chain 3-like protein 2 antibody
    • MAP1A/1B light chain 3 A antibody
    • MAP1A/1B light chain 3 B antibody
    • MAP1A/1BLC3 antibody
    • MAP1A/MAP1B LC3 A antibody
    • MAP1A/MAP1B LC3 B antibody
    • MAP1A/MAP1B light chain 3 B antibody
    • MAP1ALC3 antibody
    • MAP1BLC3 antibody
    • MAP1LC3A antibody
    • Map1lc3b antibody
    • Microtubule associated protein 1 light chain 3 alpha antibody
    • Microtubule associated protein 1 light chain 3 beta antibody
    • Microtubule associated proteins 1A/1B light chain 3 antibody
    • Microtubule associated proteins 1A/1B light chain 3A antibody
    • Microtubule associated proteins 1A/1B light chain 3B antibody
    • Microtubule-associated protein 1 light chain 3 beta antibody
    • Microtubule-associated proteins 1A/1B light chain 3B antibody
    • MLP3B_HUMAN antibody
    see all

Images

  • Western blot - Anti-LC3A/B antibody (ab128025)
    Western blot - Anti-LC3A/B antibody (ab128025)
    All lanes : Anti-LC3A/B antibody (ab128025) at 1/1000 dilution

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : HeLa + Chloroquine Whole Cell Lysate
    Lane 3 : NIH3T3 Whole Cell Lysate
    Lane 4 : U87-MG Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 14,16 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab128025 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • Western blot - Anti-LC3A/B antibody (ab128025)
    Western blot - Anti-LC3A/B antibody (ab128025)
    All lanes : Anti-LC3A/B antibody (ab128025) at 2 µg/ml

    Lane 1 : WT
    Lane 2 : atg5-/-

    Predicted band size: 15 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-LC3A/B antibody (ab128025)
    Immunocytochemistry/ Immunofluorescence - Anti-LC3A/B antibody (ab128025)This image is courtesy of an anonymous Abreview

    ab128025 staining LC3A/B in MEF cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 10% goat serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in 10% goat serum) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Chen H  et al. MicroRNA-155 affects oxidative damage through regulating autophagy in endothelial cells. Oncol Lett 17:2237-2243 (2019). Read more (PubMed: 30675289) »
  • Zhang E  et al. Minocycline promotes cardiomyocyte mitochondrial autophagy and cardiomyocyte autophagy to prevent sepsis-induced cardiac dysfunction by Akt/mTOR signaling. Apoptosis 24:369-381 (2019). Read more (PubMed: 30756206) »
See all 35 Publications for this product

Publishing research using ab128025? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-9 of 9 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3A/B antibody

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Chinese Hamster Cell (CHO cell)
Specification
CHO cell
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 22 2015

Western blot abreview for Anti-LC3A/B antibody

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15% Gel)
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Dec 12 2014

Western blot abreview for Anti-LC3A/B antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15% Gel)
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Dec 05 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3A/B antibody

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 14 2014

Western blot abreview for Anti-LC3A/B antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (heart tube)
Specification
heart tube
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More

Abcam user community

Verified customer

Submitted Jul 14 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3A/B antibody

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Rat Cell (h9c2)
Specification
h9c2
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 08 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3A/B antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Mouse Cell (MEF)
Specification
MEF
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 08 2014

Western blot abreview for Anti-LC3A/B antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More

Abcam user community

Verified customer

Submitted Jul 08 2014

Question

Dear Technical Support Team,



A customer has enquired about Anti-LC3A/B antibody (ab128025):



“Hi



I would like some information regarding an Abcam antibody to LC3 (ab128025).



The product datasheet states that it can be used on paraffin tissue sections.



Does this product work without pretreatment such as antigen retrieval with citrate buffer pH 6.0?



Is there any reference using paraffin sections and immunohistochemistry that you can forward to me?



Hope to hear from someone soon.”





Kind regards,

Read More
Answer

Thank you for your inquiry.

We do not know of any references where this antibody was used in IHC-P. But I would like to reassure your customer that ab128025 is tested and guaranteed for IHC-P.

Please find the protocol that used to test ab128025 at the end of the email.

I hiope this information is helpful and wish you and your customer a good weekend.



Not antigen retrieval was used.
1. Fixation and Sectioning
a. Fix dissected tissues in 2% paraformaldehyde, Bouin's solution, or other fixative for
30 minutes to overnight. (NOTE: Fixation time will depend on species, tissue origin
and size of tissue sections).
b. Embed tissue in paraffin.
c. Section the tissues 5-10 micrometers thick.
d. If using parafomaldehyde as fixative antigen retrieval may be necessary.
2. Deparaffinization and Tissue Rehydration
a. Deparaffin samples by incubating sections 2-3 times in xylene for 10 minutes each.
b. Hydrate samples by placing twice in 100% ethanol for 3 minutes each, then in
95%, 70%, 50%, 30% ethanol for 2 minutes each.
3. Blocking and Primary Antibody Incubation
a. Block slides with 10% serum from the species from which the secondary antibody
was taken or 8% BSA.
b. Incubate for 30 minutes to 1 hour at room temperature in a humidified chamber.
c. Wash samples in PBS containing 2% BSA for 5 minutes.
d. Incubate slides in a humidified chamber overnight with primary antibody solution
(primary antibody in PBS containing 2% BSA).
e. Wash three times with PBS containing 2% BSA for 5 minutes.
4. Secondary Antibody Incubation and Detection
a. Incubate slides in a humidified chamber with secondary antibody solution
(secondary antibody in PBS containing 2% BSA) according to manufacturer's
protocols.
b. Wash three times with PBS containing 2% BSA for 5 minutes.
c. Detect according to manufacturer's protocols.

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