Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free (AB283660)

This data was developed using ab283650, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HCT 116 cells labelling Id1 with ab283650 at 1/50 (9.64 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in HCT 116 cell line. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free (AB283660)

This data was developed using ab283650, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (Human colorectal carcinoma epithelial cell) cells labelling Id1 with ab283650 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG (DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free (AB283660)

This data was developed using ab283650, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight observed is consistent with what has been described in the literature (PMID : 11034201, PMID : 1644289).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Id1 antibody [EPR24304-18A] (<a href='/en-us/products/primary-antibodies/id1-antibody-epr24304-18a-ab283650'>ab283650</a>) at 1/1000 dilution

Lane 1:

MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 16 kDa

Observed band size: 14-16 kDa

false

• WB

Lab

Western blot - Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free (AB283660)

This data was developed using ab283650, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a higher sensitivity ECL substrate.

In materials from rats, ab283650 is suitable only for WB application.

Exposure time : 92 seconds

All lanes:

Western blot - Anti-Id1 antibody [EPR24304-18A] (<a href='/en-us/products/primary-antibodies/id1-antibody-epr24304-18a-ab283650'>ab283650</a>) at 1/1000 dilution

Lane 1:

Untreated PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 2:

PC-12 treated with 50 ng/ml EGF (<a href='/en-us/products/proteins-peptides/recombinant-human-egf-protein-animal-free-ab9697'>ab9697</a>) for 48 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 16 kDa

Observed band size: 14-16 kDa

false

• WB

Lab

Western blot - Anti-Id1 antibody [EPR24304-18A] - BSA and Azide free (AB283660)

This data was developed using ab283650, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 8 seconds

All lanes:

Western blot - Anti-Id1 antibody [EPR24304-18A] (<a href='/en-us/products/primary-antibodies/id1-antibody-epr24304-18a-ab283650'>ab283650</a>) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with human Id1 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 16 kDa

Observed band size: 14-16 kDa

false