Immunofluorescence Customer Review for Anti-β-tubulin Antibody (STJ97037)

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Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain. TUBB3 plays a critical role in proper axon guidance and mantainance. Anti-β-tubulin -http://www.stjohnslabs.com/b-tubulin-antibody-p-98672 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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ANTIBODY CUSTOMER REVIEW
Findings: “Some background staining seen on
the negative control but very good staining
otherwise.” – Marie Davies, The University of
Sheffield
Knowledge Dock Business Centre, University Way, London E16 2RD
T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: info@stjohnslabs.com w: www.stjohnslabs.com
Protocol
Tissue Processing When cells were cultured to
70%-80% plating density on
coverslips, cells were fixed in
4% PFA for 20 mins at room
temperature before application
of antibodies and plating
coverslips on slides with DAPI
Fluoromount.
Primary
Antibody
Probing
Primary antibody was added to the
blocking solution at 1:100 for 1hr
room temperature.
Fixing 4% PFA in PBS 20 mins room
temperature
Washing 1X PBS wash for 3 times.
Permeabilization 1X PBS 0.4% Triton-X-100 15
mins room temperature
Secondary
Antibody
Probing
Secondary antibody was diluted
1:500 and DAPI was diluted 1:1000
in 1% bovine serum albumin in PBS
and incubated for 1 hr at room
temperature.
Washing 1X PBS wash for 3 times. Washing 1X PBS wash for 3 times
Blocking 1X PBS 0.01% Triton-X-100 0.2%
fish skin gelatin 1hr room
temperature
Mounting Mounted coverslips on slides with
DAPI fluoromount. Observed on
Confocal Microscopy
Model Number STJ97037
Antibody Name Anti-β-tubulin antibody
Application Immunofluorescence
Species Human
Tissue NSC-34 Cells
Dilution 1:100
Secondary Antibody Anti-mouse IgG Alexa
Fluor 488
Secondary Dilution 1:2000
Fig. In NSC-34 cell line, there was
detection of B-tubulin by STJ97037
Actin-B-tubulin Antibody (Figure:
Left : No Primary Antibody Staining,
Right: STJ97037 Actin-B-tubulin
Antibody 1:100)

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