Product Datasheet
Profilin 1 Antibody - BSA Free
NBP1-19344
Unit Size: 0.1 ml
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Publications: 1
Protocols, Publications, Related Products, Reviews, Research Tools and Images at:www.novusbio.com/NBP1-19344
Updated 12/19/2023 v.20.1

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Page 1 of 4 v.20.1 Updated 12/19/2023

NBP1-19344
Profilin 1 Antibody - BSA Free
Product Information
Unit Size

0.1 mlConcentration

1.17 mg/ml

Storage
Clonality

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw
cycles.
Polyclonal

Preservative

0.1% Sodium Azide

Isotype

IgG

PurityImmunogen affinity purified

Buffer

PBS and 30% Glycerol

Target Molecular Weight

14 kDaProduct Description
HostRabbit

Gene ID

5216

Gene Symbol

PFN1

Species

Human, Mouse, Rat, Bovine

ImmunogenSynthetic peptide made to a C-terminal portion of human Profilin 1 (withinresidues 100-140). [Swiss-Prot# P07737]

Product Application Details
Applications
Recommended Dilutions
Application Notes

Western Blot, Simple Western, Immunohistochemistry, ImmunohistochemistryParaffin
Western Blot 0.5 ug/ml, Simple Western 1:200, Immunohistochemistry 1:100,Immunohistochemistry-Paraffin 1:100
This Profilin 1 antibody is useful for Immunohistochemistry on formalin fixedparaffin embedded sections and Western blot, where a band is seen at approx.
14 kDa.
In Simple Western only 10 - 15 uL of the recommended dilution is used per datapoint. Separated by Size-Wes, Sally Sue/Peggy Sue.

Page 2 of 4 v.20.1 Updated 12/19/2023

ImagesWestern Blot: Profilin 1 Antibody [NBP1-19344] - Western Blot on HeLa
whole cell extracts.

Immunohistochemistry: Profilin 1 Antibody [NBP1-19344] - Staining ofProfilin in mouse thymus.

Simple Western: Profilin 1 Antibody [NBP1-19344] - Simple Westernlane view shows a specific band for Profilin 1 in 0.5 mg/ml of HeLa
lysate. This experiment was performed under reducing conditions using
the 12-230 kDa separation system.

Publications
van Blitterswijk M, Baker MC, Bieniek KF et al. Profilin-1 mutations are rare in patients with amyotrophic lateralsclerosis and frontotemporal dementia. Amyotroph Lateral Scler Frontotemporal Degener 2013-05-02 [PMID:23634771] (IHC-P, Human)

Page 3 of 4 v.20.1 Updated 12/19/2023

Procedures
Western Blot protocol for Profilin 1 Antibody (NBP1-19344)
Profilin 1 Antibody:Western Blot Protocol
1. Perform SDS-PAGE (4-12% MES) on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transferapparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer ofproteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations
and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10
minutes each.
7. Dilute the rabbit anti-Profilin primary antibody (NBP1-19344) in blocking buffer and incubate 1 hour at roomtemperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10
minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturersinstructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated asrequired to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).
**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%,
provided it does not interfere with antibody-antigen binding.

Immunohistochemistry-Paraffin protocol for Profilin 1 Antibody (NBP1-19344)
Profilin 1 Antibody:
Immunohistochemistry-paraffin embedded sections
Antigen Unmasking
Bring slides to a boil in 10 mM sodium citrate buffer pH 6.0 then maintain at a sub-boiling temperature for 10 minutes.
Cool slides on bench top for 30 minutes.
Staining
1. Wash sections in dH2O three times for 5 minutes each.
2. Wash section in wash buffer (1X PBS/0.1% Tween-20 (1X PBST)) for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1X PBST, 5% goat serum) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul primary antibody diluted in 1X PBST, 5% goat serum to eachsection. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated secondary antibody, diluted in 1X PBST, 5% goat serum. Incubate 30 minutes at roomtemperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Striptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in dH2O.
12. Counterstain sections in hematoxylin.
13. Wash sections in dH2O two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

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This product is for research use only and is not approved for use in humans or in clinical diagnosis.
Primary Antibodies are guaranteed for 1 year from date of receipt.
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