Antibodies, Proteins, Kits and Reagents for Life Science

318 288 ₸ Product size

100 µl 318 288 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ALKBH5 antibody [EPR18958] (ab195377)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18958] to ALKBH5
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-ALKBH5 antibody [EPR18958]
See all ALKBH5 primary antibodies
Description

Rabbit monoclonal [EPR18958] to ALKBH5
Host species

Rabbit
Specificity

ab195377 has been tested in IHC-P on Human, Mouse and Rat testis sections. It worked well on Human, but not on the Mouse or Rat sections, under our experimental conditions.

Tested Applications & Species

Application	Species
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Wild type MEF, HeLa, HepG2 , PC-12, NIH/3T3, Jurkat, Neuro-2a, HEK-293T and F9 whole cell lysates. IHC-P: Human testis and kidney tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18958
Isotype

IgG
Research areas

Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

All lanes : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : ALKBH5 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 44 kDa
Observed band size: 48 kDa
why is the actual band size different from the predicted?

Lanes 1 - 2: Merged signal (red and green). Green - ab195377 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab195377 was shown to react with ALKBH5 in wild-type HEK-293T cells in western blot with loss of signal observed in ALKBH5 knockout cell line ab266762 (ALKBH5 knockout cell lysate ab257349). Wild-type and ALKBH5 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab195377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human testis (PMID: 21264265; PMID: 27590511; PMID: 17979886) is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human kidney (PMID: 21264265) is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

Lanes 1-2 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution
Lanes 3-5 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/2000 dilution

Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
Lane 5 : F9 (mouse embryonic testicular cancer cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 44 kDa
Observed band size: 35-44 kDa why is the actual band size different from the predicted?

Exposure time : Lanes 1 and 2: 15 seconds; Lanes 3 and 4: 26 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular mass observed is consistent with the literature (PMID 21264265).
Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

All lanes : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/5000 dilution

Lane 1 : Wild type MEF whole cell lysate
Lane 2 : ALKBH5 knockout MEF whole cell lysate
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 44 kDa
Observed band size: 35-44 kDa why is the actual band size different from the predicted?

Exposure time: 70 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The Wt and ALKBH5 KO cell lysates were kindly provided by an anonymous collaborator.

The molecular mass observed is consistent with the literature (PMID 21264265).
Anti-ALKBH5 antibody [EPR18958] (ab195377)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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