Overview

  • Product name

  • Description

    Rabbit polyclonal to Talin 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken
  • Immunogen

    Synthetic peptide corresponding to Human Talin 1 aa 1650-1750 (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab71332)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: Jurkat, Ramos, MEF1 ICC/IF: HeLa cell line

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab71333 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 Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 270 kDa (predicted molecular weight: 270 kDa).Can be blocked with Human Talin 1 peptide (ab71332).
Flow Cyt Use a concentration of 3 - 4 µg/ml.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

Images

  • All lanes : Anti-Talin 1 antibody (ab71333) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 270 kDa
    Observed band size: 270 kDa
    Additional bands at: 100 kDa, 230 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes
  • IHC image of Talin 1 staining in Human Normal Kidney FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab71333, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse endoderm cell

    Preparation:

    Fix in 3% PFA in PBS for 30 min at RT

    Primary antibody: Rabbit anti-talin (ab71333), 1:100

    Secondary antibody: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy3® 488) preadsorbed (ab97075), 1:100

    Nuclear counterstain: DAPI

    Rhodamine-phalloidin, 1:100

    Nuclei were counterstained with DAPI

  • ab71333 stained in HeLa cells. Cells were fixed with 4% paraformaldehyde (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab71333 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • FlowCyt data showing mouse platelet cells stained for Talin 1 using ab71333 (3.3µg/ml). The cells were fixed in paraformaldehyde and permeabilized with 0.1% soponin. The cells were then incubated with primary antibody (3.3µg/ml) for 20 mins. The cells were then incubated with Goat polyclonal to anti-rabbit IgG conjugated to Dylight-649.

    See Abreview

  • All lanes : Anti-Talin 1 antibody (ab71333) at 1/1000 dilution

    Lane 1 : Human platelets
    Lane 2 : Mouse platelets

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to rabbit IgG conjugated to IRDye 800CW (undiluted)

    Performed under reducing conditions.

    Predicted band size: 270 kDa


    Exposure time: 1 minute


    Detection method: Licor System

    See Abreview

References

This product has been referenced in:

  • Gao J  et al. Long noncoding RNA LINC00488 functions as a ceRNA to regulate hepatocellular carcinoma cell growth and angiogenesis through miR-330-5. Dig Liver Dis N/A:N/A (2019). Read more (PubMed: 31005556) »
  • Liang Y  et al. Downregulation of Dock1 and Elmo1 suppresses the migration and invasion of triple-negative breast cancer epithelial cells through the RhoA/Rac1 pathway. Oncol Lett 16:3481-3488 (2018). Read more (PubMed: 30127952) »
See all 12 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 15 2017

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - other (Platelet)
Specification
Platelet
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Khon Huynh

Verified customer

Submitted Jul 17 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (platelet)
Loading amount
20 µg
Specification
platelet
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 08 2012

Application
Flow Cytometry
Sample
Mouse Cell (platelet)
Specification
platelet
Preparation
Cell harvesting/tissue preparation method: fixed, washed platelets from whole blood
Sample buffer: walsh buffer
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.1% saponin
Gating Strategy
settings for platelet population, ungated

Abcam user community

Verified customer

Submitted Sep 21 2012

Answer

Thanks you for confirming the details.

The protocol looks fine to me so I am sorry we are unable to determine why the antibody not able to detect right size band? Perhaps you can try following steps for troubleshooting
- 70-80% cells confluency
- Heating lysates at 100C for 10 minutes or heating at 60-70C for 15-20 minutes could be tried.
- Try no primary control
- or finally as per your suggestion different gel can also be tried.

I hope these suggestions will help. Should this antibody fails again please let me know I will replace the antibody.

Read More

Question

Product code: 71333
Lot number: 931321
Inquiry: Hello, I have had troubles with the specificity of the Talin-1 Antibody (ab71333, purchased ). Conditions and protocol as follows: - Loaded 20ug protein - SDS-PAGE was used, a standard running buffer was used, run at 120V for ˜2 hours. - Transblot Turbo from Bio-Rad was used for transfer. This used standard semi-dry consumables following the protocol set out in the Transblot manual. The time of transfer was reduced to 20 min as we had previously found that 30 min lead to proteins being transfered to the filter paper. - Blocking buffer was 5% BSA in TBST for 1 hour at 4 degrees - Primary antibody was at 1ug/ml (recommended) in blocking buffer, incubated overnight (˜18hours) at 4 degrees - detected using fresh HRP-conjugated secondaries (known to be working) at a concentration of 1/3000 in blocking buffer for 1 hr at 4 degrees - 3x 5 min washes with TBST occurred between primary and secondary incubations as well as after secondary incubation. Membranes were kept in TBS after final wash. In the sample image uploaded the first lane (juxtaposed on the image) is a MW standards ladder, followed by 4 lanes of RWPE1 cell lysate, and 4 lanes of PC3 cell lysate. There is a very faint band at just above 250kDa, which is possibly Talin-1, however there are multiple bands further down the membrane which are much darker (for example: ˜60kDa, ˜40kDa and ˜35kDa). This appears to be a specificity issue as the 270kDa band should be the most defined band. Other factors (eg concentration of antibodies) appear to be fine as other bands are showing up. Any suggestions would be welcome.

Read More
Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- Which lysis buffer was used? Was it ice cold whencells lysis was done?
-Did you try any sort of protocol troubleshooting?
- Was the lysate heated at 100C for 5 minutes?
- Could you please re-attach the image?
- Have you added any protease inhibitor in lysis buffer?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More

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