Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker is a rabbit recombinant monoclonal antibody that is used to detect alpha Tubulin in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Drosophila melanogaster, Human, Mouse, Pig, Rat samples.
- Specificity confirmed with THY1 knockout cell line validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 230 publications
- Trusted since 2007
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Tested | Tested | Expected | Expected |
Rat | Tested | Tested | Expected | Expected |
Drosophila melanogaster | Expected | Expected | Expected | Expected |
Pig | Tested | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Pig | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/50000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/50000 | Notes - |
Species Pig | Dilution info 1/1000 - 1/50000 | Notes - |
Species Human | Dilution info 1/1000 - 1/50000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info 1/1000 - 1/50000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/250 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info - | Notes - |
Species Pig | Dilution info - | Notes - |
Species Drosophila melanogaster | Dilution info 1/250 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 - 1/50 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info - | Notes - |
Species Pig | Dilution info - | Notes - |
Species Drosophila melanogaster | Dilution info 1/20 - 1/50 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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Tubulin is the major constituent of microtubules, a cylinder consisting of laterally associated linear protofilaments composed of alpha- and beta-tubulin heterodimers. Microtubules grow by the addition of GTP-tubulin dimers to the microtubule end, where a stabilizing cap forms. Below the cap, tubulin dimers are in GDP-bound state, owing to GTPase activity of alpha-tubulin.
TUBA1, TUBA4A, Tubulin alpha-4A chain, Alpha-tubulin 1, Testis-specific alpha-tubulin, Tubulin H2-alpha, Tubulin alpha-1 chain
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker is a rabbit recombinant monoclonal antibody that is used to detect alpha Tubulin in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Drosophila melanogaster, Human, Mouse, Pig, Rat samples.
- Specificity confirmed with THY1 knockout cell line validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 230 publications
- Trusted since 2007
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
This antibody is expected to recognise most alpha tubulin proteins and not only TUBA4A.
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, WB in drosophila melanogaster, human, mouse, pig, rat samples.
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) has been cited over 240 times in peer reviewed journals and is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Anti-TUBA4A antibody) (ab52866) has 28 independent reviews from customers.
alpha Tubulin antibodies are often used as loading controls in Western Blot. Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker has been verified in Western Blot samples and detects a band at 50kDa Molecular weight.
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) specifically detects alpha Tubulin (UniProt ID: P68366; Molecular weight: 50kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).
Conjugation-ready, carrier free format available for antibody clone EP1332Y - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free ab216650.
Antibody clone EP1332Y is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 594, PE, Alexa Fluor® 555 (Alexa Fluor® 488 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab185031, Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573, Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272, PE Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab208752, Alexa Fluor® 555 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab275113).
Alpha Tubulin (TUBA4A) is a fundamental component of microtubules, which are crucial for maintaining the cytoskeleton and facilitating cell division. Abnormalities in alpha Tubulin are linked to neurodegenerative diseases such as Alzheimer's Disease, Parkinson's Disease, Amyotrophic Lateral Sclerosis, and Huntington's Disease, as well as cancer, highlighting its disease relevance. Post-translational modifications of alpha Tubulin play a significant role in cellular functions and disease mechanisms. Our TUBA4A antibody is a valuable tool for studying these pathways and understanding the molecular basis of these conditions.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Alpha tubulin also known as DM1A is an important component of the structural unit called microtubules. This protein weighs approximately 50 kDa and acts as a building block in the cytoskeleton. Alpha tubulin is expressed in various cell types across many organisms playing a fundamental role in maintaining cell shape and enabling intracellular transport. Researchers often acetylate alpha tubulin to study post-translational modifications and its effects on microtubule dynamics.
Alpha tubulin combines with beta tubulin to form the tubulin dimer which is the basic unit of microtubule polymerization. These dimers assemble into microtubule filaments integral in numerous cellular processes. As part of the cytoskeleton network alpha tubulin facilitates roles in mitosis and meiosis providing spindle structures necessary for chromosome separation. Its participation in cell signaling pathways is noteworthy as it interacts with motor proteins like kinesins and dyneins to aid cellular activities.
Alpha tubulin holds critical roles in microtubule-associated processes within the cytoskeleton and intracellular transport pathways. It interacts with the MAP (microtubule-associated protein) family which modulates microtubule stability and impacts cell division and trafficking. Moreover the microtubule pathway involves proteins like tau which bind to stabilize microtubules and are implicated in cellular dynamics and signaling.
Alpha tubulin is relevant in neurodegenerative diseases such as Alzheimer's disease and cancer. Aberrant modifications or mutations in alpha tubulin or associated proteins like tau can disrupt microtubule stability contributing to pathological conditions. In cancer altered expression of alpha tubulin affects cell division leading to tumor progression and chemoresistance. Researchers study alpha tubulin using techniques such as immunofluorescence with Alexa fluor 555 to understand its role in disease better.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
The negative controls are as follows:
1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution.
2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/400 dilution.
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
The association of AJAP1 with microtubules in HUVECs is lost upon microtubule destruction. Treatment with 12.5 μM nocodazole for 24 h shows destruction of the microtubule network and loss of AJAP1 tubular localization. For a negative control, HUVECs are treated with DMSO for 24 h. Cell nuclei were counterstained with DAPI (cyan). Microscope: Zeiss LSM 780; objective lens: 63×/1.40 oil; scale bar: 25 μm.
Incubated overnight at 4°C with ab52866.
(From Figure 3E of Hotte et al)
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/10000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates at 15 µg
Lane 2: PC-12 (Rat adrenal gland heochromocytoma) whole cell lysates at 15 µg
Lane 3: NIH/3T3( Mouse embryonic fibroblast)whole cell lysates at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa
Exposure time: 1s
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab52866 staining alpha Tubulin in 293 Human embryonic kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 2 hours at 23°C. Samples were incubated with primary antibody (1/200 in 0.5% saponin) for 2 hours at 23°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-alpha Tubulin antibody
Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling alpha Tubulin with ab52866 at followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human stomach.The section was incubated with Anti-DLL3 antibody [EPR22592-18] ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Alpha Tubulin Flow Cytometry (Intracellular) staining using rabbit Anti-alpha Tubulin antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell)cells labelling alpha Tubulin with ab52866 at 1/2000 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/5000 dilution
Lane 1: Mouse brain lysates at 10 µg
Lane 2: C6 (Rat glial tumor cell line) whole cell lysates at 10 µg
Lane 3: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg
Lane 4: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysates at 10 µg
Lane 5: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates at 10 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa
Alpha Tubulin Flow Cytometry (Intracellular) staining using rabbit Anti-alpha Tubulin antibody
Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed HepG2 (human liver hepatocellular carcinoma cell line) cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/20000 dilution
Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 50 kDa
Alpha Tubulin Western blot staining of Rat brain lysates using rabbit Anti-alpha Tubulin antibody
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/50000 dilution
All lanes: Rat brain lysates at 10 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-alpha Tubulin antibody
Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-alpha Tubulin antibody
Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-alpha Tubulin antibody
Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Alpha Tubulin Western blot staining of Human fetal kidney lysates using rabbit Anti-alpha Tubulin antibody
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/50000 dilution
All lanes: Human fetal kidney lysates at 10 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52866 overnight at 4°C. Antibody binding was detected using Anti-Rabbit Alexa Fluor® 790 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/1000 dilution
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg
Lane 3: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 4: Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 5: NIH/3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
Lane 6: PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at 1/10000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa
Alpha Tubulin Western blot staining of Pig skeletal muscle lysates using rabbit Anti-alpha Tubulin antibody
All lanes: Western blot - Anti-alpha Tubulin antibody [EP1332Y] - Loading Control (ab52866) at 1/5000 dilution
All lanes: Pig skeletal muscle lysates at 20 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-alpha Tubulin antibody
Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
False colour image of Western blot: Anti-PVRIG/CD112R antibody staining at 1 ug/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red.In Western blot, Anti-PVRIG/CD112R antibody ab168153 was found to be non-specific.
A band was observed at 38 kDa in wild-type Jurkat cell lysates with no change observed in the PVRIG knockout cell line Human PVRIG knockout Jurkat cell line ab274900 (knockout cell lysate Human PVRIG knockout Jurkat cell lysate ab274958). To generate this image, wild-type and PVRIG knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane.
Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
All lanes: Western blot - Anti-PVRIG/CD112R antibody (Anti-PVRIG/CD112R antibody ab168153) at 1 µg/mL
Lane 1: Wild-type Jurkat cell lysate at 20 µg
Lane 2: PVRIG knockout Jurkat cell lysate at 20 µg
Lane 3: Daudi cell lysate at 20 µg
Lane 4: Daudi cell lysate at 10 µg
Lane 5: HeLa cell lysate at 20 µg
Lanes 1 - 5: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution
Lanes 1 - 5: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Anti-THY1 antibody [7E1B11] (Anti-CD90 / Thy1 antibody [7E1B11] ab181469) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD90 / Thy1 antibody [7E1B11] ab181469 was shown to bind specifically to THY1. A band was observed at 25-37 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in THY1 knockout cell line Human THY1 (CD90) knockout U-2 OS cell line ab262490 (knockout cell lysate Human THY1 (CD90) knockout U-2 OS cell lysate ab263925). To generate this image, wild-type and THY1 knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CD90 / Thy1 antibody [7E1B11] (Anti-CD90 / Thy1 antibody [7E1B11] ab181469) at 1/1000 dilution
Lanes 1 - 5: Western blot at 20 µg
Lane 2: Western blot - Human THY1 (CD90) knockout U-2 OS cell lysate (Human THY1 (CD90) knockout U-2 OS cell lysate ab263925)
Lane 2: Western blot - Human THY1 (CD90) knockout U-2 OS cell line (Human THY1 (CD90) knockout U-2 OS cell line ab262490)
All lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 17 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
False colour image of Western blot: Anti-MAGEA4 antibody [OTI1F9] staining at 1/4000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-MAGEA4 antibody [OTI1F9] ab139297 was shown to bind specifically to MAGEA4. A band was observed at 40 kDa in wild-type A431 cell lysates with no signal observed at this size in MAGEA4 knockout cell line Human MAGEA4 knockout A-431 cell line ab273857 (knockout cell lysate ab273811). To generate this image, wild-type and MAGEA4 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-MAGEA4 antibody [OTI1F9] (Anti-MAGEA4 antibody [OTI1F9] ab139297) at 1/4000 dilution
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: Western blot - Human MAGEA4 knockout A-431 cell lysate (ab273811) at 20 µg
Lane 3: U-2 OS cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
All lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 40 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Western blot: Anti-SEMA4D antibody [30/CD100] (Anti-Semaphorin 4D/CD100 antibody [30/CD100] ab307685) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Semaphorin 4D/CD100 antibody [30/CD100] ab307685 was shown to bind specifically to SEMA4D. A band was observed at 90-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in SEMA4D knockout cell line. To generate this image, wild-type and SEMA4D knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Semaphorin 4D/CD100 antibody [30/CD100] (Anti-Semaphorin 4D/CD100 antibody [30/CD100] ab307685) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human SEMA4D knockout HCT116 cell line (Human SEMA4D knockout HCT116 cell line ab289144)
Lane 2: SEMA4D knockout HCT 116 cell lysate at 20 µg
Lane 3: HDLM-2 UNBOILED cell lysate at 20 µg
Lane 4: PC-3 UNBOILED cell lysate at 20 µg
Lanes 1 - 4: Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 90-120 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Western blot: Anti-MGMT antibody [MT3.1] (Anti-MGMT antibody [MT3.1] ab39253) staining at 1/500 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-MGMT antibody [MT3.1] ab39253 was shown to bind specifically to MGMT. A band was observed at 22 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MGMT knockout cell line Human MGMT knockout HCT116 cell line ab286541. The band observed in the knockout lysate lane below 22 kDa is likely to represent a truncated form of MGMT. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MGMT knockout HCT 116 cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-MGMT antibody [MT3.1] (Anti-MGMT antibody [MT3.1] ab39253) at 1/500 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human MGMT knockout HCT116 cell line (Human MGMT knockout HCT116 cell line ab286541)
Lane 2: MGMT knockout HCT 116 cell lysate at 20 µg
Lanes 1 - 2: Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2: Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 22 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Western blot: anti-ACK antibody staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin (ab52866) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 118 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in TNK2 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes: Western blot at 1/1000 dilution
Lane 1: Wild-type HCT 116 at 20 µg
Lane 2: Western blot - Human TNK2 knockout HCT116 cell line (ab301274) at 20 µg
Lane 3: HeLa at 20 µg
Lane 4: PC-3
All lanes: Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
Performed under reducing conditions.
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Western blot: Mouse Monoclonal[2D2] to ARID1B Anti-ARID1B antibody [2D2] ab57461 staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin (ab52866) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 200 kDa in Wild-type HCT 116 Nuclear cell lysates with no signal observed at this size in ARID1B knockout HCT 116 Nuclear cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes: Western blot - Anti-ARID1B antibody [2D2] (Anti-ARID1B antibody [2D2] ab57461) at 1/1000 dilution
Lane 1: Wild-type HCT 116 Nuclear at 20 µg
Lane 2: Western blot - Human ARID1B knockout HCT116 cell line (Human ARID1B knockout HCT116 cell line ab287219) at 20 µg
Lane 3: RKO at 20 µg
Lane 4: PC-3 Membrane at 20 µg
All lanes: Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 244 kDa
Observed band size: 200 kDa
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
Western blot: TIGAR Antibody (E-2): sc-166290 absc-166290 staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin (ab52866) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 30 kDa in Wild-type A549 cell lysates with no signal observed at this size in TIGAR knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes: Western blot at 1/1000 dilution
Lane 1: Wild-type A549 at 20 µg
Lane 2: TIGAR knockout A549 at 20 µg
Lane 3: HepG2 at 20 µg
Lane 4: SK-OV-3 at 20 µg
All lanes: Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
Performed under reducing conditions.
Alpha Tubulin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of Human kidney using rabbit Anti-alpha Tubulin antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human kidney staining alpha Tubulin with ab52866 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab52866 anti- alpha Tubulin [EP1332Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
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