Anti-RACGAP1/MGCRACGAP antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

Paraffin embedded human testis tissue stained for RACGAP1/MGCRACGAP using ab2270 at 4 μg/ml in immunohistochemical analysis.

• ICC

Supplier Data

Immunocytochemistry - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

Immunoctochemistry analysis of paraformaldehyde fixed MCF7 (human breast adenocarcinoma cell line) cells labeling RACGAP1/MGCRACGAP with ab2270 at 10 μg/mL. Cells permeabilized with 0.15% Triton. Primary incubation 1 hour followed by Alexa Fluor^® 488 secondary antibody (4 μg/mL). The nuclear stain is DAPI (blue). Negative control : Unimmunized goat IgG (10 μg/mL) followed by Alexa Fluor^® 488 secondary antibody (4 μg/mL).

• WB

Supplier Data

Western blot - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

Primary incubation was 1 hour. Detected by chemiluminescence.

All lanes:

Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270) at 1 µg/mL

Lane 1:

K562 cell lysate (in RIPA buffer) at 35 µg

Lane 2:

Jurkat cell lysate (in RIPA buffer) at 35 µg

Predicted band size: 71 kDa

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• WB

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Western blot - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

ab2270 staining (0.2μg/ml) of Human Testis lysate (RIPA buffer, 30μg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

All lanes:

Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270) at 0.2 µg/mL

All lanes:

Human Testis lysate (RIPA buffer) at 30 µg

Predicted band size: 71 kDa

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• WB

Unknown

Western blot - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

All lanes:

Western blot - Anti-RACGAP1/MGCRACGAP antibody (ab2270) at 1 µg/mL

All lanes:

K562 cell lysate at 35 µg

Predicted band size: 71 kDa

Observed band size: 75 kDa

true

• WB

CiteAb

Western blot - Anti-RACGAP1/MGCRACGAP antibody (AB2270)

RACGAP1/MGCRACGAP western blot using anti-RACGAP1/MGCRACGAP antibody ab2270. Publication image and figure legend from Taulet, N., Vitre, B., et al., 2017, Nat Commun, PubMed 29203870.

ab2270 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab2270 please see the product overview.

IFT proteins associate with central spindle MTs in anaphase and are required for their organization. a Proximity ligation assay (PLA) showing the association between α-tubulin and IFT88 in anaphase LLC-PK1 cells. α-tubulin and IFT88 antibodies were used alone as negative controls. Maximum projections of PLA staining (left), α-tubulin (α-tub-FITC) middle panel and DAPI (inset) are shown. Magnifications, right : single confocal section of the boxed area at the central spindle. b Western blots (left) showing the amount of IFT27 and IFT88 in LLC-PK1 cells transfected with control (CT), IFT27 or IFT88 siRNA. α-tubulin : loading control. Immunofluorescence images (right) showing a decrease of IFT88 staining in an anaphase cell upon IFT88 depletion. c Immunofluorescence images of anaphase GFP-α-tubulin LLC-PK1 showing abnormal central spindle MTs organization in IFT27 and IFT88-depleted cells. Insets : DAPI. Maximum projections are shown. Magnifications, right : single confocal section of the dashed boxes. d Percentage of anaphase cells with abnormal central spindle MTs organization. n > 50 cells, 3 experiments. Mean + /- s.d. ns : not significant compared to non-treated (NT) condition and ***p < 0.001 compared to non-treated and control (t test). e Images from time-lapse microscopy showing the mitotic progression of GFP-αtubulin LLC-PK1 cells transfected with CT or IFT88 siRNA. Time (min). f Depletion of endogenous IFT27 (pig siRNA) is rescued by the expression of mCherry-IFT27 (human cDNA) resistant to IFT27 pig siRNA. Immunofluorescence images (top) showing central spindle MTs organization in LLC-PK1 cells depleted in IFT27 and expressing or not mCherry-IFT27. Graph (bottom) : percentage of anaphase cells with MTs defects. n > 30 cells, 3 experiments. Mean + /- s.d. ns : not significant and **p < 0.01 compared to mCh-IFT27 negative cells (t test). g Immunofluorescence images of anaphase LLC-PK1 cells showing defects in MKLP1 and MgcRacGAP localization. α-tubulin/MKLP1/DAPI (left panel) and α-tubulin/MgcRacGAP/DAPI (right panel) stainings are shown. h Percentage of cells with MKLP1 or MgcRacGAP localization defects in anaphase. n > 50 cells. 3 experiments. Mean +/- s.d. **p < 0.01 and ***p < 0.001 compared to control (t test). i Western blots showing equal amounts of MKLP1 and MgcRacGAP in LLC-PK1 cells transfected with the indicated siRNA. α-tubulin : loading control. Scale bars : 10 μm for main images, 3 μm for insets

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